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Correlations between clinical symptoms, leukocyte sensitivity, antigen-binding capacity, and Prausnitz-Küstner activity in a longitudinal study of ragweed pollinosis
Correlations between clinical symptoms, leukocyte sensitivity, antigen-binding capacity, and Prausnitz-Kiistner activity in a longitudinal study of ragweed pollinosis Howard L. Melam, M.D., Jacob J. Pruzansky, Roy Patterson, M.D.,” Chicago, Ill.
Ph.D., and
a six-year study of a ragweed-sensitive individ~l, a longitudinal approach ~a.3 wed to correlate symptoms of potliwsti, leukocyte reactivity to antigen, serum antigen-binding capacity, and levels of serum reaginic a&body. Fluctuation of In
symptoms paralleled changes in leukocyte sensitivity to ragweed antigen. A decrease in the patient’s reaginio antibody titer to ragweed occwrred follozving treatment ad persisted after treatment was stopped. Antigen-binding activity of the patient’s sera increased with treatment bzlt decreased after treatment was stopped. The antibody responsible for antigen-binding adivity in this patient was localized to the immunoglobulin G 01a.w.
D
espite the long use of antigen injection therapy for pollinosis, explanations for clinical efficacy are not complete. Studies have shown that with immunotherapy changes may occur in the degree of peripheral leukocyte sensitivity to antigen as measured by histamine release. I-6 Increases in the serum antigenbinding capacity or level of serum blocking antibody have also been observed after immunotherapy.l-’ A third parameter, altered following injection therapy, is the decrease in reaginic antibody reported by Connell and Sherman8 The present study was a six-year longitudinal evaluation of a ragweedsensitive individual, in which correlation of symptoms of pollinosis, peripheral
From the Allergy-Immunology Section of the Department of Medicine and the Department of Microbiology, Northwestern University Medical School. Supported in part by United States P&k Health Service Training Grant 5 TO1 AI00057-10 and Research Grant 2 ROl AI-06139-06 from the National Institute of Allergy and Infectious Disease, and in part by the Ernest 5. Bazley Asthma Research Fund to Chicago Wesley Memorial Hospital, Chicago, Ill. Received for publication March 19, 1970. *Ernest 8. Bazley Professor Allergy and Immunology.
292
Volume Number
46 5
blood leukocyte sensitivity to antigen, serum antigen-binding capacity, and levels of serum reaginic antibody was attempted. These data are described in detail because we believe that an intensive study of individual cases may provide information regarding the mechanisms responsible for effective immunotherapy. CLINICAL
SUMMARY
The patient was a 34-year-old male native of Iran, who moved to the United States in 1954 and developed ragweed rhinitis in 1961. In 1963, a diagnosis of severe ragweed rhinitis and asthma was made on the basis of history, physical examination, and positioc cutaneous tests to ragweed antigen. Ragweed immunotherapy was started at that time. Immunotherapy was stopped by the patient, without medical approval, because of the absence of symptoms during the ragweed season of 1964. He had received 80,000 protein nitrogen units (PNU) of ragweed antigen. Minor symptoms were experienced during the ragweed pollen season in 1965 and progressively severe symptoms returned during the ragweed seasons of 1966 and 1967. Treatment was resumed in September, 1967. Immunotherapy was stopped by the patient in December, 1968, after he had received a total of 205,000 PNU of ragweed antigen and had been asymptomatic during the ragweed season of 1968. He had no symptoms during the 1969 ragweed season.
MATERIALS AND METHODS Antigen and isotope labeling and
Purified labeled
Binding
ragweed fraction AIV-C was with 1311 as previously reported.10
of
‘3’l-labeled
ragweed
prepared
(RWI
by
the
method
of
King
and
Normana
antigen
Increasing amounts of 1311 RW were added to aliquots of serum. The 13x1 RW bound to human gamma globulin was coprecipitated by addition of an excess of rabbit antiserum against human Cohn fraction II. Radioactivity was determined in the precipitates and supernatants. Curves were constructed by plotting the percentage of antigen in the precipitate against the quantity of antigen added. The quantity of antigen bound by 1 ml. of each serum in slight antigen excess was determined.
Autoradiographic
immunoelectrophoresis
Immunoeleotrophoresis of preparations were thoroughly incubated for 48 hours at 4” Kodak Ready-Pak No Screen to localize the immunoglobulins
Histamine
release
from
serum was done with rabbit antihuman serum.11 After washed, the troughs were charged with 1311-labeled RW C. The slides were washed, dried, and placed in contact x-ray film. The slides were then stained with thiazine which bound labeled antigen.
the and with red
leukocytes
Leukocytes were obtained from the patient and washed. The reactivity of peripheral leukocytes as measured by histamine release was determined with the technique previously reported.7 Release of greater than 30 per cent of cellular histamine in response to a challenge with antigen is considered significant.3
RESULTS Histamine
release
from
leukocytes
The pattern of washed leukocyte histamine release during the patient’s course is shown in Table I. Before treatment 1CF3 pg of ragweed antigen E induced release of 59 per cent histamine from peripheral leukocytes. After the
59
63
68
72
8-68
12-687
5-69
S-69
I course
course
9 months
5 months
2nd
5 months
1st
therapy
80,000
68,000
54,000
40,000
18,000
6,000
II
serum
posttherapy
in slight
II
antigen
course
excess.
N.D.
N.D.
N.D.
posttherapy
N.D.
205,000
17
14
12
1
N.D.
course
I
N.D.
18
17
16
19
N.D.
N.D.
17
N.A.
10
29
19
19 29
N.D.
16
13 N.D.
29
N.D.
60
60
N.D.
0.76
4.80
N.D.
N.D.
1:250
1:250
N.D.
1:250
1: 1,000
1 :l,OOO 8.32
6.99
N.A. 1:2,000
0.48
N.A.
N.A.
N.A.
N.A.
N.A.
21
1.07
N.A.
24
44
N.A.
N-411
N.A.
0.012
N.A.
<
N.A.
1
N.A.
75
59
adaeat
release
and
19
N.D.
79
N.D.
43
40
N.D.
N.D.
22
27
54
79
65
20
N.D.
N.D.
19
18
22
63
8
59
25
antigen
N.D.5
78,000
started
course
started
33,000
therapy
dose*
125,000
of
posttherapy
of
Cwnulative
*Expressed as PNU of crude ragweed extract. tPer 5 ml. aliquot of leukocytes. SExpressed as cg RW bound by 1 ml. of whole $N.D. = not done. 1jN.A. = serum not available. TTherapy discontinued.
57
6-68
12
Q-64u
53
10
7-64
2-68
8
5-64
17
6
3-64
48
3
12-63
2-65
2
S-67
0
Month
S-63
I
11-63
Date
histamine
from leukocytes
cent
capacities,
.ug RW
1 Per
I. Histamine release from leukocytes, serum antigen-binding passive transfer titers during the patient’s course
Table
Volume 46 Number 5
Longitudinal
stud~j of ragweed pollinosis
2%
patient received 40,006 PNU of ragweed antigen, 10-l pg of ragweed antigen induced an insignificant release of histamine. The leukocyte sensitivity had decreased more than 100-fold during this period of treatment. Five months after treatment was stopped, a significant release of histamine occurred following addition of 1V2 pg of RW to the patient’s leukocytes. Four years after the first course of treatment had been stopped, the patient desired resumption of injection therapy with ragweed. During the interval after his first course of treatment, he had increasing symptoms with each succeeding ragweed .sesson. Leukocyte sensitivity had returned to the original pretreatment level since a significant quantity of histamine was released aft~er incubation with 1V3 pg of RW. After 12 months of treatment and the administration of 205,009 PNU of ragweed antigen, the patient had an asymptomatic ragweed season. Leukocyte sensitivity showed at least a lOO-fold decrease with an insignificant release of histamine to 10-l pg of RW. The patient again decided not to continue treatment after 15 months of immunotherapy and a ragweed season without symptoms. Leukocytes have continued to be nonsensitive to RW, and he has passed a second consecutive ragweed season without symptoms. Serum
binding
of
RW antigen
Serum antigen-binding capacities were done on several sera, and the binding curves are shown in Fig. 1. There was an increase in antigen binding by serum obtained following treatment when compared to the values before the first and the second courses of therapy. After immunotherapy was stopped the antigen binding of the patient’s serum decreased in May, 1969, and September, 1969. The quantity of antigen bound in slight antigen excess by 1 ml. of each
I.~G
RW
AOOEO
Fig. 1 Binding of 1311 RW by sera at different periods in the patient’s course. a, September, 21,Febmary, 1968; c, June, 1968; d, May, 1969; e, September, 1969.
1967;
296
Fig.
Melaam, Pruzansky,
J. November,
and Patterson
Allorg. 1970
2
a, Immunoelectrophoresis rabbit antihuman serum.
of pretreatment b, Autoradiograph
and posttreatment of Fig. 2, a.
sera.
Center
trough
contains
serum is shown in Table I. Before the patient’s first course of treatment, 1 ml. of the serum bound less than 0.012 pg of RW. Serum taken 3 years after the first course of treatment was stopped and just before the second course of therapy was started bound 0.48 pg of RW per milliliter. After the patient received 78,000 PNU of ragweed extract, 1 ml. of the serum bound 8.32 pg of antigen protein, a 17-fold increase. Five months after the second course of therapy was stopped, the patient’s ability to bind RW had decreased 30 per cent from its maximum capacity. The antigen-binding capacity of the patient’s serum decreased further 9 months after treatment was stopped and was only 1.5-fold greater than the level observed before the initiation of the second course of treatment. Autoradiographic
immunoelectrophoresis
Autoradiographic immunoelectrophoretic studies on pretreatment serum and serum 12 months after treatment are shown in Fig. 2. The IgG of the posttreatment serum binds the RW labeled with 1311. There is minimal binding by the pretreatment serum. Double gel diffusion autoradiographic preparations of the posttreatment serum (Fig. 3) developed with anti-alpha and anti-gamma chain sera demonstrate binding activity only by IgG of the patient’s serum. Prausnitz-Kiistner
(P-K)
titrations
Table I shows the results of P-K tests of serial sera planted on a single recipient. The highest dilution of pretreatment serum to react with ragweed challenge was a dilution of 1:2,000. Following treatment, sera diluted more than 1:250 failed to react to challenge. This reduction in P-K titer has persisted even though treatment has been stopped.
Volume Number
46 5
A
B
Fig. 3 A, Double and
gel anti-gamma
diffusion (right)
preparation chain sera.
of
posttreatment
B, Autoradiograph
serum developed of Fig. 3, A.
with
anti-alpha
(left)
DISCUSSION Fig. 4 depicts graphically the temporal relationship between the patient’s clinical symptoms and changes in his reaginic antibody titer, leukocyte reactivity to RW, and serum antigen-binding capacity. In this study an individual received two brief but intensive courses of immunotherapy for ragweed rhinitis and asthma. The seasonal symptoms of the patient were markedly reduced during certain periods of observation. These symptoms recurred one year after cessation of the first course of therapy and increased in severity until treatment was resumed. Serial studies of leukocyte sensitivity demonstrated that the decline and reappearance of symptoms paralleled similar fluctuations in leukocyte response. The results are further evidence that the in vitro leukocyte system provides a method for evaluation of immunotherapy and should help provide an explanation of one of the mechanisms of action of this type of therapy. Antibody activity in the sera of the patient, as measured by antiglobulin coprecipitation, increased during t,reatment. Nine months following ces&ion of the second course of therapy the level of antigen-binding activity had decreased U-fold from the activity during treatment. Despite this drop in antigen-binding capacity, the patient remained asymptomatic. The component of the serum responsible for the antigen-binding activity was investigated. Although immunoglobulins of the IgA and IgG classes were identified in one of the posttreatment sera of this patient, only the IgG class was shown to bind 1311labeled RW. Changes in the patient’s reaginic antibody titer were found in this study.
298
Melam, Pruzansky,
and
Patterson
h-t
1
Fig.
J. Allerg. November, 1970 -4
P
4
A, Tempora o-o; hista necessary t = Periods a Antigen bir titer (o- - -( t = Periods
sequence of symptoms and leukocyte histamine release to RW. Symptoms score, line release, o- - -0, expressed as the minimal amount of RW (micrograms) cause a significant release of histamine from a 5 ml. aliquot of leukocytes. F treatment. B, Temporal sequence of serum antigen-binding activity and P-K titers. ling (o-o) expressed as micrograms of RW bound per milliliter of serum. P-K ) expressed as the highest dilution of serum resulting in positive transfer. 3f treatment.
t
Lower levels of reaginic antibody as determined by P-K tests persisted despite discontinuation of treatment. This study shows that, at least in this patient, changes in antigen-binding capacity, leukocyte sensitivity, and reaginic antibody correlate in time sequence with symptomatic relief following injection therapy. The parallel between symptomatic improvement, decreased leukocyte sensitivity, and decreased reaginic antibody is most striking. However, other factors of equal or greater importance in immunotherapy may be discovered in the future. REFERENCES
1. Pruzansky, individuals.
J. J., and Pattersonl R.: I. Use of several antigens,
Histamine J. ALLEN.
release from leukocytes 38: 315, 1966.
of
hypersensitive
Volume Number
Longitudinal
46 5
study of ragweed pollinosis
299
release from leukocytes of hypersensitive 2. Pruzansky, J. J., and Fatterson, R.: Histamine individuals. II. Reduced sensitivity of leukocytes after injection therapy, J. ALLERG. 39: 44,1967. 3. Melam, H., Pruzansky, J. J., and Patterson, R.: Histamine release from leukocytes with antigen E and whole ragweed extract, J. ALL=. 45: 43, 1970. 4. Pruzansky, J. J., and Patterson, R. : Immunologic changes during hyposensitization therapy, J. A. M. A. 203: 805, 1968. 5. Lichtenstein. L. M.. Norman. P. S.. and Winkenwercler. W. L.: Clinical and in vitro studies on rble of i.&munothe;apy in’ragweed hay fever, Amer. J. Med. 44: 514, 1968. 6. Sadan, N., Rhyne, M. B., Mellits, E. D., Goldstein, E. O., Levy, D. A., and Lichtenstein, L. M.: ImmunotheraDv of Dollenosis in children. New Ene. J. Med. 280: 623. 1969. 7. Pruzansky, J. J., an’d PaGerson, R.: Binding ‘of Il3l-lzbeled ragweed antigen by sera of ragweed-sensitive individuals, J. ALL~G. 35: 1, 1964. 8. Conn&l. J. T.. and Sherman. , ‘W. B.: Skin-seniitizing antibodv titer. III. Relationshin of the skin-sehsitizing antibody titer to the intracutaneous &in test, to the tolerance of injections of antigens, and to the effects of prolonp.ed treatment with antigen. J. y
ALLEG.
35: 169.1964‘1
9. King, T. P., a& Norman, P. S.: Isolation studies of allergens from ragweed pollen, Biochemistry 1: 709, 1962. 10. Pruzansky, J. J., and Patterson, R.: Estimation of the homogeneity of purified ragweed antigens, J. Immun. 95: 48, 1965. 11. Crowle, A. J.: In Immunodiffusion, New York, 1961, Academic Press, Inc., p. 268,
Ph.D., and
a six-year study of a ragweed-sensitive individ~l, a longitudinal approach ~a.3 wed to correlate symptoms of potliwsti, leukocyte reactivity to antigen, serum antigen-binding capacity, and levels of serum reaginic a&body. Fluctuation of In
symptoms paralleled changes in leukocyte sensitivity to ragweed antigen. A decrease in the patient’s reaginio antibody titer to ragweed occwrred follozving treatment ad persisted after treatment was stopped. Antigen-binding activity of the patient’s sera increased with treatment bzlt decreased after treatment was stopped. The antibody responsible for antigen-binding adivity in this patient was localized to the immunoglobulin G 01a.w.
D
espite the long use of antigen injection therapy for pollinosis, explanations for clinical efficacy are not complete. Studies have shown that with immunotherapy changes may occur in the degree of peripheral leukocyte sensitivity to antigen as measured by histamine release. I-6 Increases in the serum antigenbinding capacity or level of serum blocking antibody have also been observed after immunotherapy.l-’ A third parameter, altered following injection therapy, is the decrease in reaginic antibody reported by Connell and Sherman8 The present study was a six-year longitudinal evaluation of a ragweedsensitive individual, in which correlation of symptoms of pollinosis, peripheral
From the Allergy-Immunology Section of the Department of Medicine and the Department of Microbiology, Northwestern University Medical School. Supported in part by United States P&k Health Service Training Grant 5 TO1 AI00057-10 and Research Grant 2 ROl AI-06139-06 from the National Institute of Allergy and Infectious Disease, and in part by the Ernest 5. Bazley Asthma Research Fund to Chicago Wesley Memorial Hospital, Chicago, Ill. Received for publication March 19, 1970. *Ernest 8. Bazley Professor Allergy and Immunology.
292
Volume Number
46 5
blood leukocyte sensitivity to antigen, serum antigen-binding capacity, and levels of serum reaginic antibody was attempted. These data are described in detail because we believe that an intensive study of individual cases may provide information regarding the mechanisms responsible for effective immunotherapy. CLINICAL
SUMMARY
The patient was a 34-year-old male native of Iran, who moved to the United States in 1954 and developed ragweed rhinitis in 1961. In 1963, a diagnosis of severe ragweed rhinitis and asthma was made on the basis of history, physical examination, and positioc cutaneous tests to ragweed antigen. Ragweed immunotherapy was started at that time. Immunotherapy was stopped by the patient, without medical approval, because of the absence of symptoms during the ragweed season of 1964. He had received 80,000 protein nitrogen units (PNU) of ragweed antigen. Minor symptoms were experienced during the ragweed pollen season in 1965 and progressively severe symptoms returned during the ragweed seasons of 1966 and 1967. Treatment was resumed in September, 1967. Immunotherapy was stopped by the patient in December, 1968, after he had received a total of 205,000 PNU of ragweed antigen and had been asymptomatic during the ragweed season of 1968. He had no symptoms during the 1969 ragweed season.
MATERIALS AND METHODS Antigen and isotope labeling and
Purified labeled
Binding
ragweed fraction AIV-C was with 1311 as previously reported.10
of
‘3’l-labeled
ragweed
prepared
(RWI
by
the
method
of
King
and
Normana
antigen
Increasing amounts of 1311 RW were added to aliquots of serum. The 13x1 RW bound to human gamma globulin was coprecipitated by addition of an excess of rabbit antiserum against human Cohn fraction II. Radioactivity was determined in the precipitates and supernatants. Curves were constructed by plotting the percentage of antigen in the precipitate against the quantity of antigen added. The quantity of antigen bound by 1 ml. of each serum in slight antigen excess was determined.
Autoradiographic
immunoelectrophoresis
Immunoeleotrophoresis of preparations were thoroughly incubated for 48 hours at 4” Kodak Ready-Pak No Screen to localize the immunoglobulins
Histamine
release
from
serum was done with rabbit antihuman serum.11 After washed, the troughs were charged with 1311-labeled RW C. The slides were washed, dried, and placed in contact x-ray film. The slides were then stained with thiazine which bound labeled antigen.
the and with red
leukocytes
Leukocytes were obtained from the patient and washed. The reactivity of peripheral leukocytes as measured by histamine release was determined with the technique previously reported.7 Release of greater than 30 per cent of cellular histamine in response to a challenge with antigen is considered significant.3
RESULTS Histamine
release
from
leukocytes
The pattern of washed leukocyte histamine release during the patient’s course is shown in Table I. Before treatment 1CF3 pg of ragweed antigen E induced release of 59 per cent histamine from peripheral leukocytes. After the
59
63
68
72
8-68
12-687
5-69
S-69
I course
course
9 months
5 months
2nd
5 months
1st
therapy
80,000
68,000
54,000
40,000
18,000
6,000
II
serum
posttherapy
in slight
II
antigen
course
excess.
N.D.
N.D.
N.D.
posttherapy
N.D.
205,000
17
14
12
1
N.D.
course
I
N.D.
18
17
16
19
N.D.
N.D.
17
N.A.
10
29
19
19 29
N.D.
16
13 N.D.
29
N.D.
60
60
N.D.
0.76
4.80
N.D.
N.D.
1:250
1:250
N.D.
1:250
1: 1,000
1 :l,OOO 8.32
6.99
N.A. 1:2,000
0.48
N.A.
N.A.
N.A.
N.A.
N.A.
21
1.07
N.A.
24
44
N.A.
N-411
N.A.
0.012
N.A.
<
N.A.
1
N.A.
75
59
adaeat
release
and
19
N.D.
79
N.D.
43
40
N.D.
N.D.
22
27
54
79
65
20
N.D.
N.D.
19
18
22
63
8
59
25
antigen
N.D.5
78,000
started
course
started
33,000
therapy
dose*
125,000
of
posttherapy
of
Cwnulative
*Expressed as PNU of crude ragweed extract. tPer 5 ml. aliquot of leukocytes. SExpressed as cg RW bound by 1 ml. of whole $N.D. = not done. 1jN.A. = serum not available. TTherapy discontinued.
57
6-68
12
Q-64u
53
10
7-64
2-68
8
5-64
17
6
3-64
48
3
12-63
2-65
2
S-67
0
Month
S-63
I
11-63
Date
histamine
from leukocytes
cent
capacities,
.ug RW
1 Per
I. Histamine release from leukocytes, serum antigen-binding passive transfer titers during the patient’s course
Table
Volume 46 Number 5
Longitudinal
stud~j of ragweed pollinosis
2%
patient received 40,006 PNU of ragweed antigen, 10-l pg of ragweed antigen induced an insignificant release of histamine. The leukocyte sensitivity had decreased more than 100-fold during this period of treatment. Five months after treatment was stopped, a significant release of histamine occurred following addition of 1V2 pg of RW to the patient’s leukocytes. Four years after the first course of treatment had been stopped, the patient desired resumption of injection therapy with ragweed. During the interval after his first course of treatment, he had increasing symptoms with each succeeding ragweed .sesson. Leukocyte sensitivity had returned to the original pretreatment level since a significant quantity of histamine was released aft~er incubation with 1V3 pg of RW. After 12 months of treatment and the administration of 205,009 PNU of ragweed antigen, the patient had an asymptomatic ragweed season. Leukocyte sensitivity showed at least a lOO-fold decrease with an insignificant release of histamine to 10-l pg of RW. The patient again decided not to continue treatment after 15 months of immunotherapy and a ragweed season without symptoms. Leukocytes have continued to be nonsensitive to RW, and he has passed a second consecutive ragweed season without symptoms. Serum
binding
of
RW antigen
Serum antigen-binding capacities were done on several sera, and the binding curves are shown in Fig. 1. There was an increase in antigen binding by serum obtained following treatment when compared to the values before the first and the second courses of therapy. After immunotherapy was stopped the antigen binding of the patient’s serum decreased in May, 1969, and September, 1969. The quantity of antigen bound in slight antigen excess by 1 ml. of each
I.~G
RW
AOOEO
Fig. 1 Binding of 1311 RW by sera at different periods in the patient’s course. a, September, 21,Febmary, 1968; c, June, 1968; d, May, 1969; e, September, 1969.
1967;
296
Fig.
Melaam, Pruzansky,
J. November,
and Patterson
Allorg. 1970
2
a, Immunoelectrophoresis rabbit antihuman serum.
of pretreatment b, Autoradiograph
and posttreatment of Fig. 2, a.
sera.
Center
trough
contains
serum is shown in Table I. Before the patient’s first course of treatment, 1 ml. of the serum bound less than 0.012 pg of RW. Serum taken 3 years after the first course of treatment was stopped and just before the second course of therapy was started bound 0.48 pg of RW per milliliter. After the patient received 78,000 PNU of ragweed extract, 1 ml. of the serum bound 8.32 pg of antigen protein, a 17-fold increase. Five months after the second course of therapy was stopped, the patient’s ability to bind RW had decreased 30 per cent from its maximum capacity. The antigen-binding capacity of the patient’s serum decreased further 9 months after treatment was stopped and was only 1.5-fold greater than the level observed before the initiation of the second course of treatment. Autoradiographic
immunoelectrophoresis
Autoradiographic immunoelectrophoretic studies on pretreatment serum and serum 12 months after treatment are shown in Fig. 2. The IgG of the posttreatment serum binds the RW labeled with 1311. There is minimal binding by the pretreatment serum. Double gel diffusion autoradiographic preparations of the posttreatment serum (Fig. 3) developed with anti-alpha and anti-gamma chain sera demonstrate binding activity only by IgG of the patient’s serum. Prausnitz-Kiistner
(P-K)
titrations
Table I shows the results of P-K tests of serial sera planted on a single recipient. The highest dilution of pretreatment serum to react with ragweed challenge was a dilution of 1:2,000. Following treatment, sera diluted more than 1:250 failed to react to challenge. This reduction in P-K titer has persisted even though treatment has been stopped.
Volume Number
46 5
A
B
Fig. 3 A, Double and
gel anti-gamma
diffusion (right)
preparation chain sera.
of
posttreatment
B, Autoradiograph
serum developed of Fig. 3, A.
with
anti-alpha
(left)
DISCUSSION Fig. 4 depicts graphically the temporal relationship between the patient’s clinical symptoms and changes in his reaginic antibody titer, leukocyte reactivity to RW, and serum antigen-binding capacity. In this study an individual received two brief but intensive courses of immunotherapy for ragweed rhinitis and asthma. The seasonal symptoms of the patient were markedly reduced during certain periods of observation. These symptoms recurred one year after cessation of the first course of therapy and increased in severity until treatment was resumed. Serial studies of leukocyte sensitivity demonstrated that the decline and reappearance of symptoms paralleled similar fluctuations in leukocyte response. The results are further evidence that the in vitro leukocyte system provides a method for evaluation of immunotherapy and should help provide an explanation of one of the mechanisms of action of this type of therapy. Antibody activity in the sera of the patient, as measured by antiglobulin coprecipitation, increased during t,reatment. Nine months following ces&ion of the second course of therapy the level of antigen-binding activity had decreased U-fold from the activity during treatment. Despite this drop in antigen-binding capacity, the patient remained asymptomatic. The component of the serum responsible for the antigen-binding activity was investigated. Although immunoglobulins of the IgA and IgG classes were identified in one of the posttreatment sera of this patient, only the IgG class was shown to bind 1311labeled RW. Changes in the patient’s reaginic antibody titer were found in this study.
298
Melam, Pruzansky,
and
Patterson
h-t
1
Fig.
J. Allerg. November, 1970 -4
P
4
A, Tempora o-o; hista necessary t = Periods a Antigen bir titer (o- - -( t = Periods
sequence of symptoms and leukocyte histamine release to RW. Symptoms score, line release, o- - -0, expressed as the minimal amount of RW (micrograms) cause a significant release of histamine from a 5 ml. aliquot of leukocytes. F treatment. B, Temporal sequence of serum antigen-binding activity and P-K titers. ling (o-o) expressed as micrograms of RW bound per milliliter of serum. P-K ) expressed as the highest dilution of serum resulting in positive transfer. 3f treatment.
t
Lower levels of reaginic antibody as determined by P-K tests persisted despite discontinuation of treatment. This study shows that, at least in this patient, changes in antigen-binding capacity, leukocyte sensitivity, and reaginic antibody correlate in time sequence with symptomatic relief following injection therapy. The parallel between symptomatic improvement, decreased leukocyte sensitivity, and decreased reaginic antibody is most striking. However, other factors of equal or greater importance in immunotherapy may be discovered in the future. REFERENCES
1. Pruzansky, individuals.
J. J., and Pattersonl R.: I. Use of several antigens,
Histamine J. ALLEN.
release from leukocytes 38: 315, 1966.
of
hypersensitive
Volume Number
Longitudinal
46 5
study of ragweed pollinosis
299
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