Studies on the heterophile antibodies present in infectious mononucleosis

65 Pages·02.521 MB·English

by  Markowitz, Abraham

STUDIES ON THE HETEROPHILE ANTIBODIES
PRESENT IN INFECTIOUS MONONUCLEOSIS
- A'Thesis :
Presented to
the F a cu lty o f the Graduate School
%
U n iv e rs ity o f Southern C a lifo rn ia
In p a rtia l F u lfillm e n t
of the Requirem ents fo r the Degree
M aster of Science
C
by
Abraham Markowitz-
June 195°
UMI Number: EP55012
All rights reserved
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This thesis, written by
ABBAEAM MARKOWITZ
under the guidance of h.X.B.. Faculty Committee,
and approved by all its members, has been
presented to and accepted by the Council on
Graduate Study and Research in partial fulfill­
ment of the requirements for the degree of
MASTER OF__SOIENOE
..............
Faculty Committee
ACKNOWLEDGEMENT
The author is indebted to Dr. Wayne L.' Simmonds,
Department of M edical M icro biolo gy, S chool.of M edicine,
U n iv e rs ity of Southern C a lifo rn ia , fo r in va lu a b le advice
and guidance in perform ing these stu d ie s.
A p p re cia tio n due to D r. E loise Jameson and Mr. H arry
B a rn e tt, Department of B iochem istry, U n iv e rs ity of Southern
C a lifo rn ia , fo r te c h n ic a l advice and assistance in perform ­
ing the e le c tro p h o re tic studies is a lso acknowledged.
TABLE OF CONTENTS
PAOtE
THE PROBLEM AND DEFINITIONS OF TERMS USED * . . . 1
The problem . . . . . . . . . . . . . 1
Statement o f the p r o b le m .................................................... 1
Im portance o f the s tu d y ................................ * . • 2
D e £ in itlo n e o f term s used 2
H ete rop hile . . . . . . . . . . . . . 2
Forssman a n tig e n . . • • • « • • • . . 2
M ononucleosis antigen . . . . . . . . . 3
Serum sickness a n tig e n . . . 3
A ntige n • 3
REVIEW OF THE LITERATURE * A
METHODS AND MATERIALS .......................................10
P rep ara tion o f guinea p ig kidney tissu e e x tra c t • 10
E x tra c tio n o f M ononucleosis a n tig e n from beef . .
e ryth ro cyte s ..................................... 12
T e sting fo r p u rity o f M -antigen . . . . . . . 15
E x tra c tio n o f serum sickness a n tig e n from guinea
p ig kidney ..................................... 15
T esting o f the SS-antigen fo r p u rity . . . . . 17
P rep ara tion o f Beef Therm ostable complex from
beef e ryth ro cyte s * • • 19
P rep ara tion o f a n ti-F serum . . . . . . • • 21
V
page
P rep ara tion o f an ti-she ep e ryth ro cyte serum . • • 21
P rep ara tion o f anti-S SA serum * * * . * . . . ' 2 2
P rep ara tion o f a n ti-M serum . . . . . . . . 22
Source o f in fe c tio u s m ononucleosis and serum
sickness sera . # 22
experim ental procedures and r e s u l t s ...............................................23
A bsorption a n a lysis . . . . . . . . . . . 23
In fe c tio u s m ononucleosis sera absorbed w ith
guinea p ig kidney a lc o h o lic e x tra c t . 23
C o n tro l sera absorbed w ith guinea p ig kidney
a lc o h o lic e x tra c t » • « • • • * « • • . 2k
In fe c tio u s m ononucleosis sera absorbed x/ith
varying amounts o f M -antigen . . . . . . . 2k
Serum sickness sera absorbed w ith SS-antigen
fo llo w e d by guinea p ig a lc o h o lic
e x tra c t a b so rp tio n . . • . .....................................................27
In fe c tio u s m ononucleosis sera absorbed w ith
M -antigen fo llo w e d by SS-antigen a b so rp tio n . . 29
E le c tro p h o re tic a n a ly s is . . . . . . . . . J2
Comparison o f hem olysin and a g g lu tin in tite r s
o f in fe c tio u s m ononucleosis sera . . . . . . 33
DISCUSSION . . . . . . . . . . . . . . . k9
SUMMARY . . . . . . . . . . . . . . . 5k
"BIBLIOGRAPHY.................................................................................. 55
LIST OF TABLES
table PAG®
I. E x tra c tio n o f M -antigen from beef stroma . . . Ik
I I . H em agglutination In h ib itio n Test of
In fe c tio u s M ononucleosis serum w ith
M -antigen . . ...................................., ........................ 16
I I I . E x tra c tio n o f SS-antigen from G-uinea p ig
Kidney T is s u e ........................................................ 18
IV . Complement F ix a tio n Test of SS-antigen w ith
a n ti-F serum ......................................... . . . . . . 20
V. In fe c tio u s M ononucleosis sera Absorbed w ith
hot a lc o h o lic G-uinea p ig Kidney e x tra c t . . 25
V I. C ontrol sera absorbed w ith G-uinea p ig kidney
Tissue e x tra c t ..................... . . . . . . . . . . 26
V II. In fe c tio u s M ononucleosis sera absorbed w ith
Varying amounts o f M -antigen .............................. . 28
V III. Serum sickness sera Absorbed w ith SS-antigen
and fo llo w e d by G-uinea p ig Kidney e x tra c t
ab sorp tion ............................................................................ 30
IX . In fe c tio u s M ononucleosis sera absorbed w ith
M -antigen and fo llo w e d by SS-antigen
ab sorp tion . . . . . . .............................................. 31
vii
tabus PAG-E
X. Percentage of Serum P ro te in F ra ctio n s as
Determ ined E le c tro p h o re tic a lly , ..................... 3A
XI. Comparison of Hem olysin and H em agglutinin
T ite rs of In fe c tio u s Mononucleosis sera * . A8
LIST OF FIGURES
FIGURE PAGE
i. Unabsorbed serum #3A, Ascending side • • * * 35
i i . Unabsorbed serum #3A# Descending side. * • . 36
i i i . Absorbed serum #3^, Ascending side. . . . . 37
iv . Absorbed serum #3^, Descending side . . • • 38
v. Unabsorbed serum #36, Ascending side . . . . 39
v i. Unabsorbed serum #3&> Descending sid e . . . . AO
v ii. Absorbed serum #36, Ascending side. . . . • A l
v iii. Absorbed serum #36, Descending side . . • A2
ix . Unabsorbed serum #3A, Ascending and
Descending sides . . . . . . . . . . A 3
x. Absorbed serum #3A, Ascending and Descending
sides . . • * • • • • • . . # * AA
x i* Unabsorbed serum #36, Ascending and
Descending sides * • • • • . . . ♦ A5
x ii. Absorbed serum #36> Ascending and Descending
sides . A6
THE PROBLEM AND DEFINITIONS OF TERMS USED
The antibody response in the disease of in fe c tio u s
m ononucleosis hag been considered to be n o n sp e cific in th a t
these a n tib o d ie s can be p a r tia lly absorbed by guinea p ig
kidney e x tra c t and presum ably com pletely absorbed w ith beef
e ryth ro cyte s. The la te r hag been considered to be o f diag­
n o s tic s ig n ific a n c e in in fe c tio u s m ononucleosis. This
procedure gives no adequate in d ic a tio n of the antibody e n tity
or e n titie s present in in fe c tio u s mononucleosis serum as to
kin d or number*
I t has been shown by absorption o f the a n tib o d ie s
present in in fe c tio u s m ononucleosis w ith guinea p ig kidney
tissu e th a t there is a drop in the t it e r which does not ex­
ceed one fo u rth of the o rig in a l t it e r . This decrease in
t it e r is held by some to be o f no s ta tis tic a l s ig n ific a n c e .
THE PROBLEM
Statement o f the problem * I t was the purpose of th is
study ( 1) to s e ro lo g ic a lly determ ine the type or types of
h e te ro p h ile a n tib o d ie s present in in fe c tio u s m ononucleosis;
(2) to present an explanation fo r the drop in t it e r during
a b so rp tio n o f such sera w ith guinea p ig kidney tis s u e ; and
(3) to dem onstrate the re la tio n s h ip between the hem olytic
and a g g lu tin a tio n tite r s in in fe c tio u s m ononucleosis sera.