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RT PCR Protocols 1st Edition Nicola King Digital Instant
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Author(s): Nicola King, Joe OConnell
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Methods in Molecular Biology TM

VOLUME 193

RT-PCR
Protocols
Edited by
Joe O’Connell

HUMANA PRESS
RT-PCR Protocols
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M E T H O D S I N M O L E C U L A R B I O L O G Y™

RT-PCR
Protocols

Edited by

Joe O’Connell
Department of Medicine,
National University of Ireland,
Cork, Ireland

Humana Press Totowa, New Jersey

©2002 Humana Press Inc.
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Cover illustration: Background from Fig. 3A in Chapter 17 “RT-PCR-Based Approaches to Generate Probes
for mRNA Detection by In Situ Hybridization” by Joe O’Connell; foreground from Fig. 2 in Chapter 18
“Amplified RNA for Gene Array Hybridizations” by Valentina I. Shustova and Stephen J. Meltzer.

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RT-PCR Protocols/edited by Joseph O’Connell

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ISBN 0-89603-875-0 (alk. paper)
1. Polymerase chain reaction--Laboratory manuals. I. O’Connell, Joseph. II. Methods in
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QP606.D46 R8 2002
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2002190221
Preface
Until the mid 1980s, the detection and quantification of a specific mRNA
was a difficult task, usually only undertaken by a skilled molecular biologist.
With the advent of PCR, it became possible to amplify specific mRNA, after
first converting the mRNA to cDNA via reverse transcriptase. The arrival of
this technique—termed reverse transcription-PCR (RT-PCR)—meant that
mRNA suddenly became amenable to rapid and sensitive analysis, without
the need for advanced training in molecular biology. This new accessibility of
mRNA, which has been facilitated by the rapid accumulation of sequence data
for human mRNAs, means that every biomedical researcher can now include
measurement of specific mRNA expression as a routine component of his/her
research plans.
In view of the ubiquity of the use of standard RT-PCR, the main objective
of RT-PCR Protocols is essentially to provide novel, useful applications of
RT-PCR. These include some useful adaptations and applications that could
be relevant to the wider research community who are already familiar with the
basic RT-PCR protocol. For example, a variety of different adaptations are
described that have been employed to obtain quantitative data from RT-PCR.
Quantitative RT-PCR provides the ability to accurately measure changes/imbal-
ances in specific mRNA expression between normal and diseased tissues.
Because of its remarkable sensitivity, RT-PCR enables the detection of low-abun-
dance mRNAs even at the level of individual cells. RT-PCR has afforded many
opportunities in diagnostics, allowing sensitive detection of RNA viruses such as
HIV and HCV. RT-PCR facilitates many diverse techniques in research, includ-
ing in situ localization of mRNA, antibody engineering, and cDNA cloning. In
particular, the present work highlights how RT-PCR complements other tech-
nological advances, such as laser-capture microdissection (LCM), real-time
PCR, microarray technology, HPLC, and time-resolved fluorimetry.
RT-PCR has become one of the most widely applied techniques in bio-
medical research, and has been a major boon to the molecular investigation of
disease pathogenesis. Determination of the pathogenesis of diseases at the
molecular level is already beginning to inform the design of new therapeutic
strategies. It is our hope that RT-PCR Protocols will stimulate the reader to
explore diverse new ways in which this remarkable technique can facilitate
the molecular aspects of their biomedical research.
Joe O’Connell

v
Contents
Preface ............................................................................................................. v
Contributors ..................................................................................................... xi

PART I. INTRODUCTION
1 RT-PCR in Biomedicine: Opportunities Arising from the New
Accessibility of mRNA
Joe O’Connell ......................................................................................... 3
2 The Basics of RT-PCR: Some Practical Considerations
Joe O’Connell ....................................................................................... 19
PART II. HIGHLY SENSITIVE DETECTION AND ANALYSIS OF MRNA

3 Using the Quantitative Competitive RT-PCR Technique

to Analyze Minute Amounts of Different mRNAs in Small
Tissue Samples
Susanne Greber-Platzer, Brigitte Balcz,
Christine Fleischmann, and Gert Lubec ..................................... 29
4 Detection of mRNA Expression and Alternative Splicing
in a Single Cell
Tsutomu Kumazaki .............................................................................. 59
5 Nested RT-PCR: Sensitivity Controls are Essential
to Determine the Biological Significance of Detected mRNA
Triona Goode, Wen-Zhe Ho, Terry O’Connor,
Sandra Busteed, Steven D. Douglas, Fergus Shanahan,
and Joe O’Connell .......................................................................... 65
PART III. QUANTITATIVE RT-PCR
6 Quantitative RT-PCR: A Review of Current Methodologies
Caroline Joyce ..................................................................................... 83
7 Rapid Development of a Quantitative-Competitive (qc)
RT-PCR Assay Using a Composite Primer Approach
Joe O’Connell, Aileen Houston, Raymond Kelly,
Darren O’Brien, Aideen Ryan, Michael W. Bennett,
and Kenneth Nally .......................................................................... 93
8 Quantitation of Gene Expression by RT-PCR and HPLC
Analysis of PCR Products
Franz Bachmair, Christian G. Huber,
and Guenter Daxenbichler ............................................................ 103

vii
viii Contents

9 Time-Resolved Fluorometric Detection of Cytokine mRNAs

Amplified by RT-PCR
Kaisa Nieminen, Markus Halminen, Matti Waris, Mika Mäkelä,
Johannes Savolainen, Minna Sjöroos, and Jorma Ilonen ...... 117
10 Mimic-Based RT-PCR Quantitation of Substance P mRNA
in Human Mononuclear Phagocytes and Lymphocytes
Jian-Ping Lai, Steven D. Douglas, and Wen-Zhe Ho ..................... 129
PART IV. DETECTION AND ANALYSIS OF RNA VIRUSES
11 Detection and Quantification of the Hepatitis C Viral Genome
Liam J. Fanning ................................................................................. 151
12 Semi-Quantitative Detection of Hepatitis C Virus RNA
by "Real-Time" RT-PCR
Joerg F. Schlaak ................................................................................ 161
13 RT-PCR for the Assessment of Genetically Heterogenous
Populations of the Hepatitis C Virus
Brian Mullan, Liam J. Fanning, Fergus Shanahan,
and Daniel G. Sullivan ................................................................. 171
PART V. IN SITU LOCALIZATION OF MRNA EXPRESSION
14 In Situ Immuno-PCR: A Newly Developed Method for Highly
Sensitive Antigen Detection In Situ
Yi Cao .................................................................................................. 191
15 RT-PCR from Laser-Capture Microdissected Samples
Tatjana Crnogorac-Jurcevic, Torsten O. Nielsen,
and Nick R. Lemoine .................................................................... 197
16 Mycobacterium paratuberculosis Detected by Nested PCR
in Intestinal Granulomas Isolated by LCM in Cases
of Crohn’s Disease
Paul Ryan, Simon Aarons, Michael W. Bennett, Gary Lee,
Gerald C. O’Sullivan, Joe O’Connell,
and Fergus Shanahan .................................................................. 205
17 RT-PCR-Based Approaches to Generate Probes for mRNA
Detection by In Situ Hybridization
Joe O’Connell ..................................................................................... 213
PART VI. DIFFERENTIAL MRNA EXPRESSION
18 Amplified RNA for Gene Array Hybridizations
Valentina I. Shustova and Stephen J. Meltzer .............................. 227
Contents ix

19 Semi-Quantitative Determination of Differential Gene Expression

in Primary Tumors and Matched Metastases by RT-PCR:
Comparison with Other Methods
Benno Mann and Christoph Hanski ................................................ 237
PART VII. GENETIC ANALYSIS
20 Detection of Single Nucleotide Polymorphisms Using
a Non-Isotopic RNase Cleavage Assay
Frank Waldron-Lynch, Claire Adams, Michael G. Molloy,
and Fergal O’Gara ........................................................................ 253
PART VIII. RT-PCR IN IMMUNOLOGY

21 Detection of Clonally Expanded T-Cells by RT-PCR-SSCP

and Nucleotide Sequencing of T-Cell Receptor β-CDR3 Regions
Manae Suzuki Kurokawa, Kusuki Nishioka,
and Tomohiro Kato ...................................................................... 267
22 Generation of scFv from a Phage Display Mini-Library Derived
from Tumor-Infiltrating B-Cells
Nadège Gruel, Beatrix Kotlan, Marie Beuzard,
and Jean-Luc Teillaud ................................................................. 281
23 Generation of Murine scFv Intrabodies from B-Cell Hybridomas
Chang Hoon Nam, Sandrine Moutel,
and Jean-Luc Teillaud ................................................................. 301
24 Quantitation of mRNA Levels by RT-PCR in Cells Purified
by FACS: Application to Peripheral Cannabinoid Receptors
in Leukocyte Subsets
Jean Marchand and Pierre Carayon ................................................ 329
PART IX. RT-PCR IN ANTI-SENSE TECHNOLOGY
25 Detection of Anti-Sense RNA Transcripts by Anti-Sense RT-PCR
Michael C. Yeung and Allan S. Lau ................................................. 341
PART X. RT-PCR IN CDNA CLONING
26 RT-PCR in cDNA Library Construction
Vincent Healy ..................................................................................... 349
27 An RT-PCR-Based Protocol for the Rapid Generation of Large,
Representative cDNA Libraries for Expression Screening
Joe O’Connell ..................................................................................... 363
Index ............................................................................................................ 375
Contributors
SIMON AARONS • Cork Cancer Research Centre, Mercy Hospital, Cork, Ireland
CLAIRE ADAMS • Department of Microbiology, National University of Ireland,
Cork, Ireland
FRANZ BACHMAIR • Department of Obstetrics and Gynecology, University
Hospital, University of Innsbruck, Austria
BRIGITTE BALCZ • Department of Pediatrics, Division of Pediatric Cardiology,
University of Vienna, Austria
MICHAEL W. BENNETT • Department of Medicine, National University of Ireland,
Cork, Ireland
MARIE BEUZARD • Laboratoire de Biotechnologie des Anticorps and INSERM
U255, Institut Curie, Paris, France
SANDRA BUSTEED • Department of Medicine, National University of Ireland,
Cork, Ireland
YI CAO • Division of Cellular Immunology, German Cancer Research Center,
Heidelberg, Germany
PIERRE CARAYON • Sanofi-Synthélabo, Montpellier, France
TATJANA CRNOGORAC-JURCEVIC • Molecular Oncology Unit, Faculty of Medicine,
Hammersmith Hospital, London, UK
GUENTER DAXENBICHLER • Department of Obstetrics and Gynecology, University
Hospital, University of Innsbruck, Austria
STEVEN D. DOUGLAS • Division of Immunologic and Infectious Diseases, Joseph
Stokes Jr. Research Institute at the Children’s Hospital of Philadelphia,
Philadelphia, PA
LIAM J. FANNING • Hepatitis C Unit, Department of Medicine, Cork University
Hospital, Ireland
C HRISTINE F LEISCHMANN • Division of Pediatric Cardiology, Department
of Pediatrics, University of Vienna, Austria
TRIONA GOODE • Huffington Center on Aging, Baylor College of Medicine,
Houston, TX
SUSANNE GREBER-PLATZER • Division of Pediatric Cardiology, Department
of Pediatrics, University of Vienna, Austria
NADÈGE GRUEL • Laboratoire de Biotechnologie des Anticorps, Institut Curie,
Paris, France

xi
xii Contributors

MARKUS HALMINEN • Department of Virology, University of Turku, Finland

CHRISTOPH HANSKI • Department of Gastroenterology, Universitätsklinikum
Benjamin Franklin, Freie Universität Berlin, Germany
VINCENT HEALY • Wellcome Trust Cellular Physiology Research Unit, Department
of Physiology, National University of Ireland, Cork, Ireland
WEN-ZHE HO • Division of Immunologic and Infectious Diseases, Joseph Stokes
Jr. Research Institute at the Children’s Hospital of Philadelphia, PA
AILEEN HOUSTON • Department of Medicine, National University of Ireland,
Cork, Ireland
CHRISTIAN G. HUBER • Institute of Analytical Chemistry and Radiochemistry,
University of Innsbruck, Austria
JORMA ILONEN • Department of Virology, University of Turku, Finland
CAROLINE JOYCE • Department of Biochemistry, University Hospital, Cork,
Ireland
TOMOHIRO KATO • Rheumatology, Immunology and Genetics Program, Institute
of Medical Science, St. Marianna University School of Medicine, Kawasaki,
Japan
RAYMOND KELLY • Department of Medicine, National University of Ireland,
Cork, Ireland
B EATRIX K OTLAN • National Institute of Hæmatology and Immunology,
Budapest, Hungary
TSUTOMU KUMAZAKI • Research Institute for Radiation Biology and Medicine,
Department of Biochemistry and Biophysics, Hiroshima University, Japan
MANAE SUZUKI KUROKAWA • Rheumatology, Immunology and Genetics Program,
Institute of Medical Science, St. Marianna University School of Medicine,
Kawasaki, Japan
JIAN-PING LAI • Division of Immunologic and Infectious Diseases, Joseph Stokes
Jr. Research Institute at the Children’s Hospital of Philadelphia, PA
ALLAN S. LAU • Department of Pediatrics, University of Hong Kong, Queen
Mary Hospital, Hong Kong
GARY LEE • Department of Pathology, Mercy Hospital, Cork, Ireland
NICK R. LEMOINE • Molecular Oncology Unit, Faculty of Medicine, Hammersmith
Hospital, London, UK
GERT LUBEC • Division of Pediatric Cardiology, Department of Pediatrics,
University of Vienna, Austria
MIKA MÄKELÄ • Department of Clinical Allergology and Pulmonary Diseases,
University of Turku, Finland
Contributors xiii

BENNO MANN • Department of Surgery, Universitätsklinikum Benjamin

Franklin, Freie Universität Berlin, Germany
JEAN MARCHAND • Sanofi-Synthélabo, Montpellier, France
STEPHEN J. MELTZER • University of Maryland School of Medicine, Baltimore, MD
MICHAEL G. MOLLOY • Department of Rheumatology, Cork University Hospital,
Ireland
SANDRINE MOUTEL • Laboratoire de Biotechnologie des Anticorps, Institut
Curie, Paris, France
BRIAN MULLAN • Hepatitis C Unit, Department of Medicine, National University
of Ireland, Cork, Ireland
KENNETH NALLY • Department of Medicine, National University of Ireland,
Cork, Ireland
CHANG HOON NAM • Laboratoire de Biotechnologie des Anticorps and INSERM
U255, Institut Curie, Paris, France
TORSTEN O. NIELSEN • Molecular Oncology Unit, Faculty of Medicine,
Hammersmith Hospital, London, UK
KAISA NIEMINEN • Department of Clinical Allergology and Pulmonary
Diseases, University of Turku, Finland
KUSUKI NISHIOKA • Rheumatology, Immunology and Genetics Program,
Institute of Medical Science, St. Marianna University School of Medicine,
Kawasaki, Japan
DARREN O’BRIEN • Department of Medicine, National University of Ireland,
Cork, Ireland
JOE O’CONNELL • Department of Medicine, National University of Ireland,
Cork, Ireland
TERRY O’CONNOR • Department of Medicine, National University of Ireland,
Cork, Ireland
FERGAL O’GARA • Department of Microbiology, National University of Ireland,
Cork, Ireland
GERALD C. O’SULLIVAN • Department of Surgery, National University of Ireland,
Cork, Ireland
AIDEEN RYAN • Department of Medicine, National University of Ireland,
Cork, Ireland
PAUL RYAN • Cork Cancer Research Centre, Mercy Hospital, Cork, Ireland
JOHANNES SAVOLAINEN • Department of Clinical Allergology and Pulmonary
Diseases, University of Turku, Finland
JOERG F. SCHLAAK • St. Mary’s Hospital, London, UK
FERGUS SHANAHAN • Department of Medicine, National University of Ireland,
Cork, Ireland
xiv Contributors

VALENTINA I. SHUSTOVA • University of Maryland School of Medicine,

Baltimore, MD
MINNA SJÖROOS • Department of Virology, University of Turku, Finland
DANIEL G. SULLIVAN • Department of Laboratory Medicine, University of
Washington, Seattle, WA
JEAN-LUC TEILLAUD • Laboratoire de Biotechnologie des Anticorps and
INSERM U255, Institut Curie, Paris, France
FRANK WALDRON-LYNCH • University of Cambridge Clinical School,
Addenbrooke’s Hospital, Cambridge, UK
MATTI WARIS • Laboratory of Biophysics, University of Turku, Finland
MICHAEL C. YEUNG • Snyder Research Foundation, Winfield, KS
RT-PCR in Biomedicine 1

I
INTRODUCTION
2 O'Connell
RT-PCR in Biomedicine 3

1
RT-PCR in Biomedicine
Opportunities Arising from the New Accessibility of mRNA

Joe O’Connell

1. Introduction
Reverse-transcriptase-polymerase chain reaction has become one of the most
widely applied techniques in biomedical research. The ease with which the
technique permits specific mRNA to be detected and quantified has been a
major asset in the molecular investigation of disease pathogenesis. Disease-
related imbalances in the expression of specific mRNAs can be sensitively and
quantitatively determined by RT-PCR. RT-PCR also offers many opportuni-
ties in diagnostics, allowing sensitive detection of RNA viruses such as Human
Immunodeficiency Virus (HIV) and Hepatitis C Virus (HCV). RT-PCR is an
integral component of many methodologies that are essential to biomedical
research, including in situ localization of mRNA, antibody engineering, and
cDNA cloning. This chapter provides an overview of some of the ways in which
RT-PCR can be utilized in biomedical science, and summarizes the importance
and applicability of the protocols described in this volume. These protocols
include some useful adaptations and applications that may have significance
for those in the wider research community who are already familiar with the
basic RT-PCR protocol. Each individual chapter in this volume contains com-
plete experimental detail for the protocols described, so that even a newcomer
to RT-PCR should be able to perform the techniques. In particular, this volume
demonstrates how RT-PCR complements other technologies, such as laser-cap-
ture microdissection (LCM), real-time PCR, microarray analysis, high-pres-
sure liquid chromatography (HPLC) and time-resolved fluorometry.

From: Methods in Molecular Biology, vol. 193: RT-PCR Protocols

Edited by: J. O'Connell © Humana Press Inc., Totowa, NJ

3
4 O'Connell

2. Highly Sensitive Detection and Analysis of mRNA

The greatest advantage of RT-PCR in the analysis of mRNA is its extraordi-
nary sensitivity. Using nested RT-PCR, mRNA can essentially be detected at
the level of single copies. Many of the chapters in this volume demonstrate the
highly sensitive detection of mRNA. In Chapter 4, nested RT-PCR is used to
analyze mRNA expression in a single cell; a single cell is lysed and placed
directly in the RT-PCR reaction. Using appropriate, intron-spanning primers,
differential mRNA splicing may also be analyzed by this technique at the
single-cell level (1). Although the technique was demonstrated using a single
cell in suspension, RT-PCR detection of mRNA in small numbers of cells in
solid tissues is also made possible by use of laser-microdissection (2) (see
Chapter 15). The capability for such sensitive detection and analysis of mRNA
is an enormous asset in many research areas, such as developmental biology.
For example, mRNA expression can now be analyzed from the earliest phases
of embryogenesis, at the level of only a few embryonic cells. Another area that
requires analysis of small local populations of cells is brain research. Under-
standing the function of the brain is one of the greatest challenges in biology
today. Research in brain biology will benefit significantly from RT-PCR; the
expression of low-abundance mRNAs can now be measured in small tissue
samples from specific areas of the brain (3,4). In Chapter 3, for example,
RT-PCR is applied to detect and quantify specific mRNA at the femto/attogram
level in minute amounts of brain tissue. Although the extraordinary sensitivity
of nested RT-PCR is a huge advantage for the detection of low-abundance
mRNA, this level of sensitivity also presents some risks in the interpretation of
results obtained using this technique. Chapter 5 highlights a caveat pertaining
to the use and interpretation of data from nested RT-PCR; unless a quantitative
approach is employed, sensitivity controls should be adopted to estimate the
level of mRNA detected by the nested RT-PCR assay. Otherwise, the amount
of detected mRNA can be overestimated (5).
3. Quantitative RT-PCR: Approaches and Applications
The sensitivity of RT-PCR makes it particularly useful for detecting low-
abundance mRNA, especially in small amounts of tissue such as biopsy speci-
mens. A disadvantage of standard RT-PCR with respect to less sensitive
techniques such as Northern blot is that it is only semi-quantitative. This is
because of the “plateau” in the kinetics of PCR product accumulation, in which
linearity in the relationship between product and initial template tapers off with
increasing cycles. Many strategies have been developed to enable quantitative
data to be obtained from RT-PCR. Some of the most commonly used
approaches are reviewed in Chapter 6, and methodologies and applications of
several of these approaches are explored in this volume.
RT-PCR in Biomedicine 5

Many quantitative RT-PCR approaches are based on competitive PCR (6).

Essentially, a control PCR template is constructed that has identical primer
sites to the target template, but has a difference—for example, in size—which
allows amplification products from this control template to be distinguished
from those of the target template. This control template is spiked in at known
concentration as an internal standard prior to amplification of the target tem-
plate. The standard will compete directly with the target template during PCR
amplification, so that if the internal competitive standard template is present in
equal amount to the target template, equivalent PCR products are obtained from
both. In practice, multiple PCR reactions (usually 5–7) are set up containing
serially increasing amounts of the internal standard. Following PCR amplifica-
tion, the equivalence point—where there is equal yield of target and competitive
standard PCR products—is determined. The number of copies of the target tem-
plate must be equivalent to the known number of competitive standard molecules
spiked into this particular reaction, enabling quantification of target molecules.
In order to perform competitive PCR, a control standard as described in the
previous paragraph must be constructed for each target mRNA to be quantified.
Several methods have been devised for this purpose, and indeed the construction
of standards is also facilitated by PCR. In Chapter 3, for example, a series of
overlapping PCRs is designed so that a small deletion of a few base-pairs (bp) is
created in the target cDNA sequence. The target is PCR-amplified in two sepa-
rate fragments, leaving an intervening region of a few bp between them. The
two fragments are annealed together via overlapping complementary regions
tagged onto the PCR primers. Thus, when the two fragments are annealed
together, the intervening region is deleted. The annealed fragments are then
PCR-amplified as a single product for use as the competitive standard. The
advantage of making such minor alterations to the target cDNA is that the stan-
dard will be almost identical to the target, so that there is not likely to be a
difference in the amplification efficiency between both. This is crucial to the
validity of competitive PCR, which depends on equal competition between
both templates. However, because of the close similarity between the PCR
products obtained from target and competitor, the high resolution of a DNA-
sequencing gel or column is required to separate the products for determina-
tion of the equivalence point.
A common approach to generating a competitive standard is to make a larger
deletion, usually of about 30%, in the target. This method permits differentia-
tion between amplification products of target and competitor on a standard
agarose gel (6). The advantage of this strategy is that once the standard has
been constructed, no deviation from the standard RT-PCR protocol is required;
the equivalence point is simply detected on a standard agarose gel. The com-
petitor is usually of sufficiently similar size and sequence composition to the
Other documents randomly have
different content
nobody knows; but from that City it was conveyed with great Pomp
in 1682. to Pisa in Tuscany, where it is still worshiped in a Church
bearing the pretended Saint’s Name[458]. According to the most
probable Opinion, Stephen governed Four Years, and about Six
Months.

Valerian, SIXTUS II. Gallienus.

Twenty-third Bishop of
Rome.
S tephen being dead, Sixtus or Xystus II. a Year of Christ 257.
Deacon of the Church of Rome, was chosen
to succeed him. As the late Dispute was not yet Dionysius of Alex-
andria interposes in
ended, Dionysius Bishop of Alexandria no sooner the famous Dispute.
heard of his Promotion, than he began to press
him with great Earnestness to relinquish the wild Pretensions of his
Predecessor, and concur with the other Bishops in restoring Peace
and Tranquillity to the Church[459]. He writ Three Letters to him on
the same Subject, whereof the last was from Dionysius and the
whole Church of Alexandria, to Sixtus and the whole Church of
Rome[460]. He writ likewise to Dionysius and Philemon, two
Presbyters of the Church of Rome, whom we have mentioned above,
and who upon Stephen’s Death seem to have abandoned his Party;
for Dionysius of Alexandria, in his first Letter to Sixtus, writes, That
these two Presbyters had been formerly of Stephen’s Opinion[461], a
plain Indication that they were not then. The Bishop of Alexandria
had at last the Satisfaction to see his pious Endeavours crowned with
Success; for we find no farther Mention of this Dispute till it was
revived by the Donatists. In what manner it Peace restored to
ended, we are no-where told; but it is manifest the Church by his
from the Writers of those Times, that the African means.
and Asiatic Bishops continued the same Practice of baptizing
Heretics, till it was condemned by the two great Councils, of Arles in
314. and of Nice in 325[462]. Whence we may well conclude, that the
Terms proposed at the Beginning of the Dispute by Dionysius and St.
Cyprian were agreed to by Sixtus, viz. That no Restraint should be
laid on the Bishops of either Side, but that every one should be
allowed to follow undisturbed which of the two Opinions he thought
most agreeable to the Scripture and to Reason. This was allowing
the Bishops to consult the Scriptures, and make use of their own
Reason, in a Point already judged and decided by the Bishop of
Rome. But the Successors of Sixtus have not been so complaisant;
for they pretend, that a blind Faith ought to be yielded to all their
Decisions as infallibly true, a blind Obedience to all their Decrees as
unquestionably holy.
But now the Persecution, which had begun some Months before
the Decease of Stephen, raged with more Violence than ever: For
Valerian having, at the Instigation of an Egyptian Valerian persecutes
Magician, changed the Kindness he once had for the Church.
the Christians into an implacable Hatred, he ordered, by a Rescript to
the Senate, all Bishops, Priests, and Deacons, to be carefully sought
for, and executed without Mercy[463]. Pursuant to Sixtus martyred.
this Order, Sixtus, who among the first fell into
the Hands of the Persecutors, was immediately either beheaded, as
we read in the Pontifical of Bucherius[464], or crucified, as we are told
by Prudentius[465]; having held the Chair only Eleven Months, and
some Days. Pontius, a Deacon of the Church of Carthage, styles him
a good and pacific Prelate[466], no doubt on account of his Conduct
quite opposite to that of his ambitious and quarrelsome
Predecessor[N8].

N8. Ruffinus published, under the Name of Sixtus Bishop of

Rome, the Book of a Pythagorean Philosopher, named Sixtus. St.
Jerom reproaches him in two Places with that Imposture, as he
styles it, supposing him to have known the Work, which he
ascribed to Pope Sixtus, not to be his[1]. St. Austin was imposed
upon among the rest; for, in his Treatise of Nature and Grace, he
 quotes that Book as the Work of Pope Sixtus; but he afterwards
owned and corrected his Mistake[2]. It was ranked by Pope
Gelasius among the Books of Heretics; so that he supposed it to
have been written by a Christian, which was more than he could
know, there not being a single Word in it whence we can argue
the Author to have believed in, or to have had any Knowlege of
Christ: and it is on this Consideration that it has been thought
unworthy of a Bishop of those times.

1. Ep. ad Ctesiph. contr. 2. Aug. l. 2. retract. c. 42.

Pelag. c. 22. & in cap. 18.
Ezech.

Gallienus, DIONYSIUS, Claudius II.

Twenty-fourth Bishop of
Rome.
S ixtus being dead, and the Christians prevented Year of Christ 258.
by the Persecution from assembling to chuse
another in his room, the See remained vacant The See vacant
almost
almost a whole Year, that is, from the 6th of a whole Year.
August 258. to the 22d of July 259. when
Dionysius, a Presbyter of the Church of Rome, whom we have
mentioned above, was elected, to the great Satisfaction of the
Faithful; for he was one of the most eminent Men of his Time both
for Piety and Learning[467]. During his Pontificate, the Goths broke
into the Empire, over-ran all Asia Minor, and, having almost utterly
destroyed the City of Cæsarea, they carried with them into Captivity
most of its Christian Inhabitants. Firmilian was then Bishop of the
Place, who had censured the Conduct of Stephen with so much
Sharpness and Acrimony; but the Remembrance of what had passed
on that unhappy Occasion had not that Effect on Dionysius’s Charity
Dionysius, which far less Provocations have had to
on many of his Successors; for he no sooner the distressed
Chris-
heard of the Distress that Church was in, than, tians of Cæsarea.
laying hold of so favourable an Opportunity to
exert his Charity, he writ a Letter to comfort them in their Calamity,
and at the same time dispatched proper Persons with large
Collections to ransom the Christians who had fallen into the Hands of
the Barbarians[468]. The Letter, which Dionysius wrote on this
Occasion, was carefully kept in the Archives of the Church of
Cæsarea, as an authentic Monument of his Goodness and
Charity[469]. The great Dionysius Bishop of Alexandria having, at this
Time, composed a learned Treatise to prove against Sabellius the
Distinction of the Divine Persons, some over-zealous Catholics,
misconstruing several Passages in that Work, and concluding that he
had run into the opposite Error, accused him to the Bishop of Rome,
as if he denied the Son to be consubstantial with the Father[470].
Hereupon the Bishop of Rome, having Dionysius of Alex-
assembled a Council, acquainted Dionysius with andria, accused at
the Sentiments of the other Bishops, and his Rome over him.
own, expressing his Concern, that the Divinity of the Word should
have been questioned by him, and at the same time desiring him to
answer the Accusation[471]. This Dionysius readily did in Four Books,
which he styled Confutation and Apology; shewing therein that his
Opinion was very different from what it had been represented at
Rome, and explaining those Passages which had given Ground for
the Accusation. This Work he addressed to the Bishop of Rome[472].
Here Baronius exults. Behold, says he, one of the most eminent
Prelates of the Church, upon Suspicion of Heresy, arraigned at Rome,
judged at Rome. Who does not see a supreme That argues no
Tribunal erected there, to which all Causes must Jurisdiction in the
be brought; a sovereign Judge residing there, by Bishop of Rome.
whom all Persons must be absolved or condemned; is either blind
and cannot see, or shuts his Eyes and will not see[473]. And does not
the sharp-sighted Annalist himself see what every one the least
conversant in Ecclesiastical History must see, if he is not either blind
and cannot, or shuts his Eyes and will not see, viz. Bishops, when
guilty, or only suspected of Heresy, accused to some of their
Collegues, who neither had nor claimed any Jurisdiction over them?
Thus was the famous Paul of Samosata, Bishop of Antioch, at this
very Time, accused by his whole Church, first to Dionysius Bishop of
Alexandria, and soon after to Firmilian Bishop of Cæsarea[474]. That
such an Accusation argued any Jurisdiction in those Bishops over the
Bishop of Antioch, is what Baronius himself dares not affirm; and yet
a like Accusation brought to Rome is enough for him to transform
that See into a supreme Tribunal; that Bishop, though far from such
ambitious Thoughts, into a sovereign Judge. But the Bishop of
Rome, says Baronius, required of Dionysius a Confession or
Declaration of his Faith: And does not that argue Superiority and
Jurisdiction? Baronius himself knew it does not: for it is impossible
he should not know, that when a Bishop was suspected of Heresy, all
his Collegues had a Right to require of him Confession of his Faith,
and not to communicate with him till they had received it.
In the Time of Dionysius was held the famous Paul Bishop of
Council of Antioch, which condemned and Antioch condemned
deposed Paul Bishop of that City, who denied the and deposed,
Distinction of the Divine Persons, and the Divinity of Christ. Of the
Deposition of Paul, and the Election of Damnus, who was placed in
his room, Notice was immediately given to the whole Church, by a
Synodal Letter addressed to Dionysius Bishop of Rome, and to
Maximus, who had succeeded the great Dionysius in the See of
Alexandria[475]. And here it will not be foreign to my Purpose to
observe, that the Bishop of Antioch was summoned to appear before
the Council, and not at the supreme Tribunal erected by Baronius at
Rome; that he was condemned and deposed without the
without the consent or Concurrence, nay, and Consent
without the Knowlege of the sovereign Judge or Knowlege of the
Bishop of Rome.
residing at Rome; that he did not appeal to him,
which he certainly would have done, as he was a Man of
unparalleled Impudence and Ambition, had such a Custom obtained
in those Days; and lastly, that the Fathers of the Council writ to the
Bishop of Rome in the same Manner as they did to other Bishops,
letting him know, that for the future he was to communicate with
Damnus, and not with Paul. All this is manifest from the Account
which St. Basil gives us of that Council[476]. And yet Baronius brings
in that Father, even on this Occasion, as an Evidence for the Papal
Supremacy[N9].

N9. For by wrong pointing a Passage in the Latin Translation of

that Author, he makes him contradict himself, and ascribe the
deposing of Paul to Dionysius Bishop of Rome, and the Great
Dionysius Bishop of Alexandria, though the latter was dead before
Paul was deposed, as is evident from the Letter which was written
by the Council on that Occasion, and is addressed to Maximus the
Successor of Dionysius in the See of Alexandria[1]. The Passage
runs thus; Duo enim Dionysii diu ante eos septuaginta fuere, qui
Samosatensem sustulere, quorum alter Romæ, alter Alexandriæ
Præsul erat[2]. The Meaning of St. Basil is, that the two Dionysius’s
flourished before the Council of Antioch, which consisted of
Seventy Bishops, and deposed Paul of Samosata; that is, before
the Second Council that was assembled against him; for another
had been convened in the same City about eight Years before to
depose him; but upon his pretending to renounce his Errors, the
Sentence had been suspended. The above-quoted Passage
Baronius stops thus; Duo enim Dionysii diu ante eos septuaginta
fuere; qui Samosatensem deposuere, &c. so that the Relative qui
refers, according to this Method of Pointing, to the Two
Dionysius’s, and not to the Seventy Bishops: as if St. Basil had
said, The Two Dionysius’s, who deposed Paul of Samosata,
flourished before the Council of Antioch, which was composed of
Seventy Bishops[3]. So that Paul must be twice deposed, St. Basil
must contradict himself, all the Writers of those Times must be
arraigned as guilty of an unpardonable Omission, lest the Bishop
of Rome should appear to have been, what he really was, an idle
Spectator of a Transaction so famous in the History of the Church.
A Writer of any Honour or Honesty had rather give up a Cause,
than expose himself thus by attempting to defend it.
 1. Euseb. l. 7. c. 30. 3. Bar. ad ann. 265. n. 10.

2. Basil. de syn. p. 918.

From St. Basil, Baronius runs to the Emperor Aurelian, begging of

a Pagan Prince what he could not extort from a Catholic Bishop, a
Declaration and Acknowlegement of the Pope’s Supremacy. The
Reader must know, that Paul having kept, by Paul keeps
Force, Possession of the Bishop’s Habitation in Possession
Defiance of the Council, the Catholic Bishops had of the Bishop’s
Habitation.
recourse to the Emperor, who, after hearing both
Parties with great Attention, adjudged the House to him, who should
be acknowleged by the Bishop of Rome, and the other Bishops of
Italy[477]. This Baronius interprets as an open Acknowlegement of the
Pope’s Supremacy; and that his Readers may not overlook it, as most
of them would be apt to do, he takes care to bespeak their
Attention, by marking it in the Margin with the following Words in
Capitals, The Emperor Aurelian acknowleges the Supremacy of the
Church of Rome[478]. From this one would expect to find Aurelian not
only turned Christian, but prostrate at his Holiness’s Feet, and
bowing down to kiss them: but our Annalist, to the great
Disappointment of his Readers, after having thus raised their
Attention, only repeats out of Eusebius the Sentence pronounced by
the Emperor, which he would have us suppose with him to have
been owing to the Knowlege that Prince had of the Pope’s
Supremacy. And why must the Pope’s The Emperor’s Sen-
Supremacy be brought in here rather than the tence whether
Supremacy of the Bishops of Ravenna, of Milan, favour-
able to the
of Aquileia, &c. and, above all, the Supremacy of Pretensions
the collective Body of the Italian Bishops? for to of the See of
them, and not to any particular Bishop, the Rome.
Cause was referred by the Emperor. As for the
Emperor’s Conduct on this Occasion, it may be thus accounted for:
That just and wise Prince observed the Bishops in the East greatly
animated against Paul; and therefore apprehending them more
sway’d by Passion and Prejudice than by Justice and Equity, he
referred the Cause to the Bishops of Italy, who, he thought, would
judge more impartially, as being placed at a Distance, and not
engaged, at least not so warmly, in the Dispute[479]. But this
happened Two Years after the Death of Dionysius dies,
Dionysius; for he died on the 26th of December
269. Claudius and Paternus being Consuls, after having governed the
Church of Rome for the Space of Ten Years, Five Months, and Four
Days, according to the most probable Opinion[480]. As he died in the
Reign of Claudius II. surnamed the Gothic, who is represented in the
Acts of some pretended Martyrs as an implacable Enemy to the
Christian Name, he is in some Martyrologies honoured with the Title
of Martyr; but as neither Eusebius, nor any other not a Martyr.
antient Writer, takes notice of that Prince’s
having ever persecuted or molested the Christians, those Acts ought
to be looked upon as fabulous, and Dionysius with 375 more
expunged out of the Catalogue of Martyrs; though some of them,
namely, Marcus, Priscus, Valentine, and Quirinus, are honoured by
the Church of Rome, as Saints of the first Class, and have filled with
their Reliques most of the Provinces of Europe.

Claudius II. FELIX, Aurelian.

Twenty-fifth Bishop of
Rome.

D
ionysius was succeeded by Felix, in whose Year of Christ 269.
Time a furious Persecution being raised by
Felix dies a Martyr
Aurelian, he may be supposed to have
in the Persecution
suffered among the rest, since he is of
distinguished by the Council of Ephesus[481], by Aurelian.
St. Cyril[482], and by Vincentius Lirinensis[483],
with the Title of Martyr. He presided, according to Eusebius[484],
Syncellus[485], and Eutychius[486], Five Years, to which Baronius adds
Eleven Months, and Twenty-five Days[487]. He writ a Letter addressed
to Maximus Bishop of Alexandria, which is quoted by Cyril, and the
Council of Ephesus[488]. The Acts of the Martyrs, who are supposed to
have suffered under Aurelian, are without all doubt supposititious;
for in them frequent Mention is made of the Emperor’s Son, whereas
the Writers of those Times tell us in express Terms, that he had a
Daughter, but no Male Issue[489].

Aurelian, EUTYCHIANUS, Probus,

Tacitus, Carus.
Twenty-sixth Bishop of
Rome.
F elix being dead, Eutychianus was chosen in his Year of Christ 275.
room in the very Beginning of the Year
275[490]. Several Things are said of him, by Eutychianus not
martyred.
Anastasius, and other Writers of no Authority;
but all I can learn of the Antients concerning him is, that he
governed Eight Years, and Eleven Months[491]; and consequently died
in the Close of the Year 283. He is honoured by the Church of Rome
as a Martyr, and said in the Roman Martyrology to have suffered
under Numerian; but it is certain that in 283. when Eutychianus died,
Numerian was not Emperor, but only Cæsar, and at that very time
engaged with his Father Carus in a War with the Persians in the East,
where he was assassinated by Aper his Father-in-Law. As for his
Brother Carinus, who remained in the West, neither he, nor the two
preceding Emperors, Tacitus and Probus, ever gave the least
Disturbance to the Christians; so that the Church of Rome must be
at the Trouble of finding out a distinct Place in Heaven from that of
the Martyrs for Eutychianus, Trophimus, Sabbacius, and the
illustrious Senator Dorymedon, who are supposed to have suffered
under those Princes.

Carus, CAIUS, Diocletian,

Carinus, Maximian.
Numerian,
Twenty-seventh Bishop
of Rome.
A s little is said by the Antients of Caius as is Year of Christ 283.V
said of his Predecessor. A few Days after the
Death of Eutychianus, Caius was chosen to succeed him, Carus and
Carinus being Consuls[492]. He presided Twelve Caius not a Martyr,
Years, Four Months, and Seven Days; that is, tho’ honoured as
from the 17th of December 283. to the 22d of a Martyr.
April 296. Caius too is counted by the Church of Rome among her
Martyrs, upon the Authority of Bede, and of the Acts of St. Susanna,
by which that Writer seems to have been misled. In those Acts Caius
is said to have suffered with Susanna his Niece, and many others,
under Numerian: but that Prince in his Father’s Life-time had no
great Power, being only Cæsar, and very young, and was killed on
his March out of Persia soon after his Father’s Death; so that he
never reigned in the West, and but a very short time in the East.
Caius therefore could not suffer under him at Rome, where his elder
Brother Carinus governed. But the Vulgar have a particular
Veneration for Martyrs, and, what turns to a very good Account, are
glad to purchase their Reliques at any rate. The The Church of
Church of Rome therefore, to provide herself Rome
with great Store of them, has multiplied beyond why so fond of
Martyrs.
Belief the Number of her Martyrs; which she
could not well do without multiplying at the same time the Number
of the Persecutors of the Christian Religion. And hence it is that
several Princes, who never molested, nay, who greatly favoured the
Christians, have been by the Church of Rome transformed in her
Martyrologies and Legends into Persecutors. As for the Acts of the
supposed St. Susanna, they are full of Mistakes and Absurdities, and
contradict the best Historians of those Times.

Dioclesian, MARCELLINUS, Constantius,

Maximian, Galerius.
Twenty-eighth Bishop
of Rome.
M arcellinus succeeded Caius on the 30th of Year of Christ 296.
June 296. and governed Eight Years, Three
Months, and Twenty-five Days, according to the Marcellinus unjustly
aspersed by the
most antient Records[493]: so that he must have Church of Rome.
died on the 24th of October 304. The Love of
Truth, which an Historian ought never to swerve from, obliges me to
undertake the Defence of this Pope against the Church of Rome
herself, and most of her Divines, who, joining the Donatists of Africa,
have endeavoured to blacken his Memory with Aspersions equally
wicked and groundless. For the Church of Rome tells us, both in her
Breviary and Martyrology, and her Divines must chime in with her,
that Marcellinus being apprehended during the Persecution of
Dioclesian, he was persuaded by that Prince to deliver up the Holy
Scripture to be burnt by the Pagans, agreeably to a late Edict, and at
the same time to offer Incense to the Gods. This they found on the
Acts of the Council of Sinuessa, which is supposed to have been
summoned on that Occasion, and before which Marcellinus is said to
have been convicted by Seventy-two Witnesses of the above-
mentioned Crimes. That such a scandalous Story, invented by the
Donatists of Africa, as St. Austin affirms[494], should not only have
been credited, but industriously propagated, by the Successors of
Marcellinus, must seem very strange and surprising to those, who
recollect with how much Zeal they have strove on other Occasions to
conceal or excuse the least Imperfections in their Predecessors. If
therefore they not only readily own the Apostasy of Marcellinus, but
are the first to divulge it, and take care to make it known in the
Breviary to those who scarce know any thing else, we may be well
assured there is a Snake hid in the Grass; the more as it is certain
almost beyond doubt, that no such Council was ever held; and
consequently that the Acts upon which alone that Apostasy is
founded, are supposititious. To unravel the Whole, the Reader must
know, that the Fall of Marcellinus made such a Noise in the Church,
as we read in those Acts, that immediately a grand Council met,
composed of no fewer than 300 Bishops. Before this Council
Marcellinus appeared; but, at his first Appearance, the Bishops,
struck with Horror at the very Thought of judging the Head of the
Church, the Judge of all, cried out with one Voice, The first See is to
be judged by nobody: accuse yourself, judge yourself, condemn
yourself. To this Testimony, so favourable to the Their View,
ambitious Views of the Bishops of Rome, is therein.
intirely owing the Sanction which they have given to such Fables,
highly injurious to the Memory of one of their best Predecessors.
Without this Lenitive the Acts of the pretended Council of Sinuessa,
supposing the Apostasy of a Pope, had been condemned; the
Absurdities and Contradictions, which it is wholly made up of, had
been set forth in a proper Light; and the Testimonies of Theodoret
and St. Austin had been alleged to vindicate the Character of
Marcellinus: for of these two Writers the former Marcellinus com-
tells us, that he acquired great Glory by his mended and
Conduct during the Persecution[495]; and the vindicated
by the Antients.
latter, in writing against Petilian the Donatist, has
the following Words: Why should I answer the Calumnies with which
he loads the Bishops of Rome? Why should I clear them from the
Crimes which he lays to their Charge? Marcellinus, and his Presbyters
Melchiades, Marcellus, and Sylvester, are accused by him as if they
had delivered up the sacred Books, and offered Incense to the Gods:
Are they therefore to be thought guilty? Does he prove what he
advances against them? He brands them with the Epithets of wicked,
and sacrilegious; but I say they are innocent: And why should I
produce Reasons to support my Defence, since he brings none to
make good his Charge[496]? But a solemn Declaration, that the See of
Rome is to be judged by nobody, made in those early Times, by 300
Bishops, carries with it such Marks of Truth, as quite invalidate the
Testimonies of Theodoret and St. Austin, and render the Apostasy of
Marcellinus, which gave room to that Declaration, undeniable! St.
Austin looks upon the Apostasy of Marcellinus, and his Presbyters
Melchiades, Marcellus, and Sylvester, who were all afterwards
Bishops of Rome, as a mere Calumny, as an Invention of the
Donatists; but their Successors, trampling upon all Authority that
stands in the Way of their Ambition, chuse rather to have Four of
their Predecessors thought Apostates and Idolaters, than part with
the Decree of that pretended Council, exalting them so high above
all other Bishops.
If Marcellinus acquired great Glory during the The Acts of the
Persecution, as Theodoret assures us; if his Council of
Apostasy was a mere Calumny, broached by the Sinuessa fabulous.
Donatists, as we read in St. Austin; the pretended Council of
Sinuessa must be given up, since it is supposed to have been
assembled on occasion of Marcellinus’s Fall: but, abstracting from the
Fall of Marcellinus, the Circumstances attending that Council are in
themselves so absurd and incredible, that there needs no other
Argument to convince a Man, who has any Understanding, and dares
to use it, that no such Council ever was, or could be held. For who
can conceive it possible, that, during the most No such Council
cruel Persecution the Church ever suffered, 300 ever held.
Bishops should assemble, not in Rome, where they might more
easily have met unobserved, but in a small Country Town, where a
much less numerous Assembly must immediately have been
observed and suspected? But, after the Death of Fabianus, says
Baronius[497], the Clergy of Rome, and the Bishops, met to chuse him
a Successor, notwithstanding the Persecution that raged then. He
ought to have said some Bishops, as St. Cyprian does[498], whom he
quotes; but I shall say so for him, that his Argument may appear in
its full Strength, and save me the Trouble of answering it; for it will
then run thus: Some Bishops, perhaps 15 or 20, met unobserved in
the great and populous City of Rome: Ergo, 300 might meet
unobserved in a small Country Town; for such was Sinuessa.
This Council is supposed to have been held in The many
a Grotto, or Cave, where there was no room but Absurdities
for 50 at a time; and yet they are all said to contained in the
Acts
have been present when Marcellinus owned his of that Council.
Crime, and divested himself of his Dignity. And
what a despicable Figure does he make on that Occasion! At first he
denies the Charge; but, being convicted by 72 Eye-witnesses, he
owns it at last, but in Terms more becoming a School-boy, trembling
at the Sight of a Rod, than a penitent Bishop, before so grave an
Assembly. But the most remarkable Passage in that Piece is the
Dispute between Urbanus High Pontiff of Jupiter, and Marcellinus
High Pontiff of the Christians. Urbanus, to convince his Fellow-Pontiff
that he ought not to scruple offering Incense to Jupiter, alleges the
Example of the Mages offering Incense to Christ. Marcellinus
answers, That the offering of Incense on that Occasion was
mysterious; and unravels the Mystery. Hereupon Urbanus,
unacquainted with Mysteries, appeals to the Judgment of the
Emperors Dioclesian and Maximian; to this Appeal Marcellinus
agrees; and the Controversy is referred by both Pontiffs to be
decided by the Two Emperors. They, no doubt, gave Sentence in
favour of Jupiter and Urbanus; and then Dioclesian, taking
Marcellinus with him into the Temple of Vesta, persuaded him there
to offer Incense to Jupiter, Hercules, and Saturn. How these Three
Deities came to have a Place in the Temple of Vesta, the Compiler of
these Acts alone knows. Such are the Absurdities and Contradictions,
of which that Piece is wholly made up. But it flatters the Ambition of
the Successors of Marcellinus; on occasion of his Fall it exalts the
See of Rome above all other Sees: its Authority therefore must not
be called in question: all the Absurdities and Contradictions it
contains, must be blindly believed; the Memory of Marcellinus most
unjustly slandered; the Testimonies of Theodoret, and St. Austin,
clearing him from all Guilt, disregarded and rejected. And may not
this be interpreted as a tacit Declaration, that they had rather he had
been guilty than innocent, provided his Guilt could any-ways
contribute to the Aggrandizing of their See? What can we think their
Ambition will spare, since they have thus sacrificed to it the
Character of one of their Predecessors, whose Memory is revered by
all Antiquity? The Church of Rome honours Marcellinus as a Saint;
and, not withstanding his pretended Apostasy, allows him a Place
amongst her Martyrs; probably by way of Reparation for the Injustice
done him. But his Martyrdom may be justly Marcellinus falsly
questioned; at least it seems to have been supposed to have
utterly unknown to St. Austin, who flourished died
a Martyr.
not long after his Time, since he never mentions
it, tho’ it would have afforded him the strongest Argument he could
possibly use to silence the Donatists. His Martyrdom, 'tis true, is
vouched by Bede, who tells us, that he was beheaded at Rome, by
Dioclesian’s Order; but that Historian is often led into gross Mistakes
by a Pontifical, supposed to have been written in the Sixth Century,
which he frequently copies, with all its Anachronisms, and other
Faults.
That, upon the Death of Marcellinus, there Vacancy of
happened a Vacancy of some Years, seems Three Years.
undeniable, since it is marked in the Pontificals, even in that of
Bucherius[499], and mentioned by all those who, till Baronius’s Time,
have written the History of the Popes: but what at this time should
occasion a Vacancy at least of Three Years, is what I will not take
upon me to account for: the Persecution lasted but Two Years in
Italy, according to Eusebius[500], which expired soon after the Death
of Marcellinus: some pretend that it raged there so long as Galerius
was Master of that Country. Be that as it will, it is certain, that
Maxentius usurped the Empire in 306. and that he not only favoured
the Christians, but pretended to be of the same Religion himself; and
yet the See remained vacant, according to the Pontifical of
Bucherius[501], till the Tenth Consulate of Maximian Hercules, and the
Seventh of Maximian Galerius, that is, till the Year 308. Baronius
indeed admits of no Vacancy; but, in Opposition to all those who
have written before him, places the Election of Marcellus immediately
after the Decease of his Predecessor Marcellinus[502]. This I should
readily agree to, but for the Authority of the above-mentioned
Pontifical, which had not yet appeared in Baronius’s Time, and is
thought to have been written about the Year 354. As for the
Chronicle of Eusebius, it can be here of no Weight on the one Side or
the other, since Marcellus is there quite left out; and his Successor
Melchiades is said to have died before Constantine made himself
Master of Rome; whereas it is certain, that, under Melchiades, a
Council was held at Rome, by that Prince’s Order, as we shall see
hereafter.

Maximian, MARCELLUS, Galerius,

Constantius, Constantine.
Twenty-ninth Bishop of
Rome.
U pon the Death of Marcellinus, the See Year of Christ 308.
remained vacant somewhat above Three
Years and an half; that is, from the 24th of October 304. to the 19th
of May 308. when Marcellus was chosen in his room. Thus says the
Pontifical of Bucherius, where, instead of Seven Years, which is a
Mistake of the Transcribers, as is manifest from the Consulships
mark’d there, we must read Three[503]. The Marcellinus and
Similitude of the Two Names has misled some Marcellus
Writers to confound Marcellinus with Marcellus; confounded
by some Writers;
for Eusebius, as well as St. Jerom, only mention
the former; and Theodoret, omitting both Marcellus and Eusebius,
who succeeded him, names Melchiades as the immediate Successor
of Marcellinus[504]; which has made Dr. Pearson doubt, whether
Marcellus was ever Bishop of Rome[505]. But but distinguished
Marcellinus and Marcellus are evidently by others.
distinguished in the Pontifical of Bucherius, by the different Times, in
which they governed, and the different Consuls, under whom their
Government began and ended[506]. They are, besides, distinguished
both by Optatus Milevitanus[507], and St. Austin[508], who speaks of
Marcellus, not only as a Presbyter of the Church of Rome, but as
Bishop of that See. To these Testimonies I may add the Epitaph of
Marcellus by Pope Damasus, supposing him to have been Bishop of
Rome[509]. Damasus flourished about the Year 366. Many things are
said of Marcellus; but they are all founded either on his Acts, or the
modern Pontificals, and consequently have no Foundation at all.
Pope Damasus, in his Epitaph, tells us, that his Steadiness in keeping
up the Discipline of the Church, and obliging such as had fallen,
during the Persecution, to give due Satisfaction, stirred up against
him a general Hatred, which, not confined to private Disputes and
Invectives, ended in Tumults, Bloodshed, and Murders[510]. Damasus
adds, that the Crime of one, who had renounced Marcellus is
the Faith, while the Church enjoyed a profound banished.
Peace, induced the Tyrant Maxentius to send Marcellus into
Banishment. But of these Transactions the Antients either have not
thought fit to give us a more particular Account, or, if they did, their
Writings have not reached our Times. Marcellus died on the 16th of
January 310. having held the Pontificate One Year, Seven Months,
and Twenty Days[511]; but whether he died in Banishment, or was
recalled to Rome, is uncertain. The Church of His Acts are
Rome, upon the Authority of his fabulous Acts, fabulous.
has added him, with many others, to the Number of her Martyrs: but
Maxentius, who reigned at Rome during his Pontificate, and under
whom he is said to have suffered, had no sooner made himself
Master of that City, than he put an End to the Persecution, as we are
told, in express Terms, by Eusebius[512]. He is His Reliques.
said to have been buried in the Cœmetery of
Priscilla, on the Salarian Way[513]: but his Body, like the Bodies of
most other Saints, is now worshiped in several Places; viz. in a
Church, bearing his Name, at Rome; in the Abbey of Omont in
Hainault, not far from Maubeuge; at Cluni, in a Parish-Church of the
Diocese of Elne in Roussillon[514], &c.
Constantine, EUSEBIUS, Licinius.
Thirtieth Bishop of
Rome.
M arcellus was succeeded by Eusebius, who Year of Christ 310.
governed Seven Months, according to
Eusebius[515], but only Four Months and Sixteen Days according to
the Pontifical of Bucherius[516]. From an antient Stands up in De-
Epitaph on this Pope we learn, that he opposed, fence of the Discip-
with great Vigour and Zeal, one Heraclius, line of the Church,
and is banished.
pretending that those who had fallen during the
Persecution, ought to be readmitted to the Communion of the
Church, without giving such Satisfaction as was then required; and
that hereupon great Divisions happening among the People,
Maxentius, to put an End to those Disturbances, banished Eusebius
into Sicily[517]. Many other things are said of him by Anastasius,
Platina, Ciacconius, and such-like Writers; but what we read in them
has no better Foundation than what is advanced by Baronius; viz.
that he instructed Eusebius the celebrated Bishop of Vercelli, and
gave him his own Name[518]; which is founded on the Acts of that
Bishop, now universally rejected as supposititious.

Constantine, MELCHIADES, Licinius.

Thirty-first Bishop of
Rome.
M elchiades, or Miltiades, as he is called in the Year of Christ 311.
antient Manuscripts, was chosen to succeed
Eusebius, on the 2d of July 311. after a Vacancy of Nine Months, and
upwards [519]; which Historians do not account for. In his Time
happened the ever memorable Conversion of Constantine con-
Constantine to the Christian Religion. That verted to the Christ-
Prince, having overcome and utterly defeated ian Religion.
the Usurper Maxentius, on the 28th of October
His Edicts in favour
312. soon after issued an Edict, jointly with of the Christians.
Licinius, who was upon the point of marrying his
Sister, allowing the Christians the free Exercise of their Religion, and
likewise the Liberty of building Churches[520]. By the same Edict he
ordered the Places, where they had held their Assemblies before the
Persecution, and which had been taken from them, to be
restored[521]. He left Rome in the Beginning of the Year 313. and,
arriving at Milan, he there issued a Second Edict, to correct some
Mistakes that had given Offence in the former[522]. What these
Mistakes were, we know not; for the Decree itself has not reached
our Times; but Valesius conjectures, that the high Commendations
bestowed on the Christian Religion alarmed the Pagans, imagining,
that the Intention of the Two Princes was to suppress theirs; and
likewise, that some Christians had taken Offence at the odious Name
of Heretics; given in that Decree to the various Sects sprung from
them[523]. Be that as it will, it is certain, that, by the Second Decree,
an intire Liberty of Conscience was granted to all sorts of Persons,
every one being allowed to honour and worship what Deity he
pleased; and in what manner soever he thought best. The Second
Edict strictly injoins all those, who had purchased of the Exchequer,
or held by Grant, any Place formerly destined for the Assemblies of
the Christians, to restore them forthwith, and apply to the
Exchequer; where they should be indemnified[524]. The same Year
313. Licinius, having gained a complete Victory over Maximinus, a
sworn Enemy to the Christians, made himself Master of Nicomedia,
and there caused the Edict of Milan to be proclaimed, and set up in
the Market-place, on the 13th of June[525]. Thus Peace was restored
to the Church, in the East as well as in the West, after a most cruel
and bloody Persecution of Ten Years, and almost Four Months; for
the First Edict against the Christians had been published in that very
City on the 24th of February 303[526]
Another remarkable Incident of this Pontificate The Schism of the
was, the famous Schism, formed in Africa Donatists in Africa.
against Cæcilianus, the Catholic Bishop of Carthage; whereof a
succinct Account will not be foreign to my Subject, as Melchiades
was chiefly concerned in most of the Transactions relating to it. The
first Decree against the Christians, published by Dioclesian, which I
have just now mentioned, ordered the Churches to be every-where
laid level with the Ground, the Books of the Scripture to be carefully
sought for, and publicly burnt; and that such Persons of Quality as
should persist in the Profession of the Christian Faith, should be
deemed infamous, and excluded from all Honours and Employments.
This Edict was executed with such Rigour in Africa, that it was a
capital Crime in the Magistrates of the Cities, and punishable with
Death, to shew any Mercy or Compassion to a Christian, who,
owning he had the sacred Books, should refuse to deliver them into
the Hands of the proper Officers. Those who, in Traditores who.
Compliance with this Edict, delivered them up,
which great Numbers did, were styled Traditores, a Name, which
afterwards became famous in the History of the Church, by affording
the Donatists a plausible Pretence to separate themselves from the
Communion of the Catholic Bishops[527]. Of this Crime Mensurius,
Bishop of Carthage, was falsly accused; but, though the Charge
could not be proved against him, yet some of his Flock, encouraged
by Donatus, Bishop of Casænigræ in Numidia, separated from his
Communion[528]. Mensurius dying some Years after, Cæcilianus,
Deacon of the Church of Carthage, was chosen in his room, in Spite
of the Cabals and Intrigues of Botrus and Cælesius, Two chief
Presbyters, who aspired to that Dignity. The chief Authors
Cæcilianus, soon after his Election, summoned of the Schism
some Persons, in whose Custody his Predecessor against Cæcilianus.
had left the Money of the Church, to deliver it up to him: but they
not only refused to comply with his Demand, but began to stir up the
People, and form a Party against him. Botrus and Cælesius were not
idle on this Occasion; but, animated with Jealousy and Envy, lest no
Art unpractised to blacken his Character, and discredit him with those
who had preferred him to them. But the chief Support of this Faction
was Lucilla, a Woman of great Quality, Wealth, and Interest, and an
avowed Enemy to Cæcilianus, who, while he was yet Deacon, had
publicly reprimanded her for kissing the Relique of a Martyr, as she
was upon the Point of receiving the Eucharist. An undeniable Proof,
that the Worship of Reliques was at this time disapproved by the
Church. Such Liberty taken with a Person of her Rank, was what she
could not brook; and therefore she laid hold of the first Opportunity
that offered, and no better could offer, to revenge the Affront[529]. It
is not to be doubted but those, who had separated from Mensurius,
joined this Faction; since the Second Schism owed its Origin to the
First, as St. Austin says, speaking of the Two Schisms under
Mensurius and Cæcilianus[530].
The Schismatics, to give an Appearance of The Bishops of
Justice and Authority to their Proceedings, Numidia summoned
summoned Secundus Bishop of Tigisis, and the to depose him.
other Bishops of Numidia, to depose Cæcilianus, and chuse another
in his room; for the Bishops of Numidia claimed the Privilege of
assisting at the Election of the Bishop of Carthage, and ordaining him
after he was elected[531]. They readily complied with the Summons;
but, upon their Arrival, they found, to their great Surprize, that the
whole City, except a small Number of Schismatics, the avowed
Enemies of Cæcilianus, communicated with him as their lawful
Bishop[532]. They were 70 in Number; but as many of them were
Traditors, and some guilty of other enormous Crimes, as appears
from the Acts of the Council of Cirtha[533], they were easily prevailed
upon by Lucilla, who is said to have spent an immense Sum on this
Occasion[534], to declare the Election of Cæcilianus void, and the See
of Carthage vacant. The only thing they could They declare his
lay to his Charge was, that he had been Election null.
ordained by Felix Bishop of Aptungus, whom they falsly accused as a
Traditor. Cæcilianus refused to Appear before them; and truly to
trust himself to such an Assembly, had been acting a very imprudent
Part; for Purpurius, Bishop of Limata, had said, If he comes among
us, instead of laying our Hands upon him, by way of Ordination, we
ought to knock out his Brains, by way of Penance[535].
The Party having thus declared Cæcilianus The Schismatics
illegally elected and ordained, they separated separate
themselves from his Communion, and from the themselves
from the
Communion of all who communicated with
him[536]; that is, from the Communion of the Communion
Catholic Church; for Cæcilianus was of the Church.
acknowleged by the other Bishops of Africa, by the Bishop of Rome,
and by all the Bishops of the World, says St. Austin[537]. Such was the
Rise of the famous Schism, which, for the Space of 300 Years, and
upwards, occasioned great Disturbances in the Churches of Africa.
Donatus, Bishop of Casænigræ in Numidia, was Called Donatists,
the first Author of it, according to St. Austin[538]; and from whom.
but it was not from him, but from Donatus, the Schismatic Bishop of
Carthage, that they took the Name of Donatists; for, till his Time,
they styled themselves the Party of Majorinus[539], whom they chose
and ordained Bishop of Carthage, in the room of Cæcilianus; though
he was then only Lector of that Church, and had been formerly one
of Lucilla’s menial Servants[540]. To justify their Conduct, and their
electing a new Bishop, they writ Letters to all the Churches of Africa,
filled with Calumnies against Cæcilianus, and those who had
ordained him. By these Letters great Numbers were imposed upon,
and misled; insomuch that, the People being every-where divided,
most Churches had Two Bishops, the one ordained by Majorinus, and
the other by Cæcilianus[541].
About this time, that is, about the Year 313. Edicts enacted by
Constantine, out of his Zeal for the Christian Constantine, in fav-
Religion, issued Two Decrees, addressed to our of the Christian
Religion.
Anulinus, Proconsul of Africa, the one
commanding all the Places in that Province to be restored, which had
once belonged to the Catholic Church, and might have been usurped
during the Persecution[542]; and the other, exempting the Ecclesiastics
from all civil Functions[543]. This Privilege was granted only to the
Ecclesiastics of the Catholic Church, whereof Cæcilianus was the
Head, as was expresly declared in the Edict; and therefore to him
alone the Proconsul imparted it. It was a great Mortification to the
Donatists to see themselves thus disregarded by the Emperor: they
therefore assembled a few Days after, and drawing up a Petition to
Constantine, they delivered it, unsealed, to Anulinus, together with a
Bundle of Papers, sealed up in a Leather Bag, with this Title: The
Petition of the Catholic Church, containing the Crimes of Cæcilianus;
by the Party of Majorinus. The Substance of the The Donatists
Petition was, that the Controversy between them petition
and the other Bishops of Africa might be referred Constantine, that
the
to the Bishops of Gaul, who were free from the Dispute may be re-
Imputation of having delivered up the sacred ferred to the
Books to the Pagans[544]. Anulinus immediately Bishops
dispatched a Messenger to the Emperor, both of Gaul.
with the Request, and the Papers, giving him, at the same time, by a
Letter still extant[545], an Insight into the Dispute, that made so great
a Noise in Africa. Constantine, who was then in Gaul, having
received and read all those Pieces, expressed great Concern to find
the Christians thus divided among themselves, and the Bishops at
Variance with one another[546]. However, he The Bishops named
readily granted to the Donatists the Judges they by Constantine.
demanded, naming, for that Purpose, Maternus Bishop of Cologne,
Rheticius Bishop of Autun, and Marinus Bishop of Arles[547]; all Men
of known Integrity, great Learning, and unblemished Characters. To
these, by a Letter under his own Hand, he gave Notice of their new
Commission; and, at the same time, for their better Information, he
caused Copies to be transmitted to them, of all the Papers he had
received from Anulinus[548]. The Three Bishops were ordered to
repair, with all Speed, to Rome, and there, jointly with Melchiades,
Bishop of that City, to sit as Judges of the Controversy. Cæcilianus
likewise was ordered to Rome, and allowed to take with him Ten
Bishops of his Party, such as he should judge the most capable of
defending his Cause; and the same Liberty was granted to the
adverse Party[549]. Constantine, in the Letter he His Letter to
writ on this Occasion to Melchiades, after Melchiades.
appealing to him as a Witness of the Respect and Veneration he had
for the Catholic Church, declares, he had nothing so much at Heart
as to see her Members happily united: he therefore earnestly intreats
him to examine the Affair with the utmost Attention, and, jointly with
the Bishops of Gaul, to judge it according to the strictest Laws of
Justice and Equity[550]. In this Letter Constantine names no other
Judges but the Three Bishops of Gaul, Melchiades, and one Mark,
supposed to have been Bishop of Milan, whom he joins with
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