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Effect of prior administration of 7S and 19S antibody on active antibody formation
Volume 65
Number 6
Part 2
here is whether or not we are dealing with primary, bilateral s y n c h r o n y - - t h a t is, a lesion in the depths in or near the midline which at times produces a diffuse abnormality and which at other times simply fires to one side as a focal spike. Obviously, I haven't the answer to this, because this would take depth recording and this would seriously impair the success of this kind of study. One can only speculate as to whether or not this is so-called secondary, bilateral synchrony, wherein a focal cortical lesion activates the subcortical pacemaker. I remind you that in 28 of our 40 families we have not been able to discover evidence for a familial abnormality. Consequently, in that group of 28 families there may be some patients who have focal structural cortical lesions. I think this certainly must be the kind of mechanism which is operating in the child with the mixed glioma in the left frontaI lobe, whose EEG you saw showing paroxysmal bilateral spike-wave discharges. In this situation one must postulate that the focal hemisphere lesion activated a subcortical pacemaker which produced the diffuse spike-wave discharges which, incidentally, are indistinguishable from pure, if there is such a thing, centrencephalic epilepsy. I think all of these findings are, among other things, a commentary on the limited repertoire of the scaIp encephalogram. DR. EDWARD F. R A ~ , 123 Garden St., Needham 92, Mass. Dr. Bray, I think this is a very thought-provoking presentation. There are two questions which come to mind. One is that some years ago there was described the phenomenon of migration of spike foci in children with age, and this observation was made as the result of the fact that some children with seizure disorder and focal spikes had surgical removal of the area of focal discharge, only with the result subsequently that the spike foci had migrated and the children sometimes persisted in having seizures. As an outgrowth of this, it was noted that children without operation also had migration of spike loci. I wondered if in any of these families that you have studied you had the opportunity to do serial EEG's over the years to find out whether the origin of these spikes that you now found in the temporal lobe began typically in the occipital lobe or in any other area. Second, I wondered if in studying so many persons you came upon any clues as to what may differentiate those persons who have spike loci seizures from those who have spike foci and no clinical manifestations of seizure disorder. DR. BRAY. Let me answer the last question first. We have no idea why some families have seizures and some do not. In a large study by Earl Walker of people who have gunshot wounds of the brain, only 35 per cent of them develop epilepsy. Obviously, the etiology of any seizure is multifactorial, there being a whole host of physiologic, biochemical factors impinging upon the nervous system to vary the state of consciousness. I think it should be said that to my knowledge there is still no convincing evidence about the basic biochemical or molecular defect in this disorder. Now, the question of migratory and shifting
Abstracts
1083
spikes has, of course, been hammered at for a long time. I think Dr. Fred Gibbs was among the first to emphasize this. Dr. Gibbs has postulated that the spike actually moves forward. He takes young infants and finds spikes in the occipital lobes which "migrate forward" on the cortex. This concept has never been very appealing to me. I would rather think that this phenomenon resuits from a slowly developing organ, the brain. The situation is very complex. For example, the neuron population is changing a great deal; the rate of myelination is changing a great deal, and, of possible importance, the temporal lobe is the last to become fully myelinated. This may be one of the reasons why a high percentage of spike discharges from the scalp over the temporal lobe is seen in adults. In order to answer Dr. Rabe's second question, I would like to show this final slide which is a graphic presentation of longitudinal EEG findings in some of these patients. You will note considerable variation in many individuals. At times their recordings will show focal epileptiform discharges. Later these abnormalities will disappear. In some cases they remain absent but in other cases, as you see, the epileptiform abnormalities will reappear again. We feel that observations of this type, based upon longitudinal studies, are important. We have called this "sampling error" which may or may not be the best term.
13. Effect o/prior administration of 7S and 19S antibody on active antibody formation Chester W. Fink and Joseph LoSpalluto, e Departments of Pediatrics, Biochemistry, and Internal Medicine, University of Texas Southwestern Medical School, Dallas, Texas In an earlier report, inhibition of antibody formation by placentaIly transmitted 7S typhoid antibody was demonstrated. In the present study the antibody response to typhoid H antigen has been examined in the rabbit following prior administration of either 7S or 19S antibody. Little or no difference between control animals receiving saline or normal rabbit serum and those receiving 19S antibody was observed. The rabbits receiving 7S antibody, however, showed both a diminished total antibody titer and a marked delay in the appearance of 7S antibody following typhoid H immunization. The differences between groups remained, even when low titers of infused 7S were compared with high titers of infused 19S antibody. Similar differences were found when typhoid H complexes in antibody excess were administered. The results indicate that the 19S antibody exerts little or no inhibitory effect on antibody formation. The specific agglutinating activities of 7S and 19S typhoid H antibody were found to be in the same order of magnitude.
1 0 84
Society for
Pedialric Research
December 1964
In addition, premature infants were immunized with typhoid-paratyphoid vaccine following exchange transfusion. Infants who had no detectable maternally transferred antibody were utilized. In 3 out of 4 infants 19S antibody acquired during exchange did not inhibit active antibody formation, whereas 7S antibody received by exchange completely inhibited it in all of 13 infants. Infants receiving no antibody, either from the mother or by exchange, responded to immunization in a normal fashion. DISCUSSION DR. HORAGE L. HODES, Mt. Sinai Hospital, 11 E. IOOth St., New York 29, N. Y. This is a very interesting paper, describing work which is very beautifully done, and we are indebted again to Dr. Fink for his contributions in this field. I think his suggestion that what we now end up with is the vestigial remains of a less efficient mechanism is a very attractive idea. The 19S antibody-forming is less than optimal in some other ways which we might mention very briefly. One is that the mechanism has a very short memory. In order to keep making 19S antibody, one must continue to receive the antigenic stimulus at short intervals. If the interval is proIonged, formation of 19S antibody stops. It is also less efficient in that 19S antibody does not diffuse so well as smaller antibodies do. I think Dr. Fink's idea that the 19S antibody mechanism is a vestigial thing is a very good one. It is possible that it had something to do with the survival of Homo sapiens as compared with the Pithecanthropus and even the later Neanderthal man, DR. FINK. Hodes.
Thank
you for your comments~
Dr.
DR. E. R. STIEHM, University o[ Cali[ornia Medical Center, San Francisco, Call[. One of the factors important in inhibiting antibody synthesis by administered antibodies would be the duration in which the antibody lasted in the system. From studies on humans, it is felt that the half-life of the 7S antibody is around 30 days and the halflife of the 19S antibody is perhaps 5 days. The half-life of rabbit gamma globulin 7S is about 7 days. I wonder if you have any information on the half-life of your 19S rabbit antibody, and if this might be a possible explanation for the failure of inhibition by the 19S a n t i b o d y - - t h a t it is just not there very long. DR. FINK. We have not done any studies on this ourselves, but there is some information in the literature. It is reported that 7S rabbit gamma globulin has a half-life of about 5 days, whereas the half-life for 19S is about 3 days. We have noted in previous and in the present experiments on humans that infants who have passively transferred 7S antibody may not produce any specific antibody for as long as 6 months after antigen administration. During at least 5 months of this time the passively transferred antibody is below detectable levels. I do not think the shorter halflife of 19S antibody explains our results, although it may be of some significance.
The h u m a n is a better subject for this type of experiment than the rabbit because of the longer half-life of the infused antibody and because the inhibition by 7S is complete rather than partial. However, it was difficult to find infants who had received exclusively 19S antibody during exchange transfusion. The few I have shown were atl that were found in three years of checking all our exchanges. DR. RICHARD T. SMITI-I, Department o/ Pediatrics, University o[ Florida, GainesviUe, Fla. Without in anyway detracting from these data, I must disagree with your interpretation and to some extent with the remarks of Dr. Hodes. Imputing a vestigial role for gamma macroglobulins does not agree with the body of data strongly supporting a major functionaI significance for macroglobulin antibodies in all higher vertebrates and mammals. Their property of preparing bacteria, effete red blood cells, and other particulate matter for phagocytosis is a role in which they appear more efficient than other classes of antibodies. They apparently can be formed when the processes which result in 7~-globulin synthesis are repressed or undeveloped. They appear very early in ontogeny. Dr. John Robbins has recently reported, on the other hand, that macroglobulins do not neutralize diphtheria toxin at all as compared with macroglobulin components directed toward the purified toxin. i n the family of immunoglobulins, each class, in all probability, has special functions and attributes. For example the YA (beta-2A) globulins are the principal secretory form of y-globulin, the 7• (macroglobulins) globulins being limited to the circulation, and the gamma-G globulins (y~) are distributed more widely in the extracellular fluid space. The assumption that macroglobuIins are vestigial and not necessary to the survival of Homo sapiens is really much too speculative to fit the data. DR. HODES. I should like to answer Dr. Smith since I was included in his discussion. The word "vestigial" applied to an organ (or to a function) does not necessarily mean that it is not useful, or that it is on its way out. Anthropologically speaking, I would suggest only that these are different mechanisms, that the 19S is earlier and associated with different phenomena. For example, the appendix is getting smaller and in that sense is vestigial. The h u m a n nose is smaI1 as compared with that of the chimpanzee, but it is still useful. It is in this sense that I used the term "vestigial." DR. FINK. I think there may be other factors that lend credence to this possibility. Phylogenic studies show that the more primitive forms have proIonged 19S production and in some the 19S form of antibody may be predominant. It is also interesting to speculate about the type of antigens which resuIt in predominantly 19S production in humans, an example of this type being typhoid O antigen. These antigens seem to be of the polysaccharide or lipopolysaccharide variety. We have done some preliminary studies in this area.
Number 6
Part 2
here is whether or not we are dealing with primary, bilateral s y n c h r o n y - - t h a t is, a lesion in the depths in or near the midline which at times produces a diffuse abnormality and which at other times simply fires to one side as a focal spike. Obviously, I haven't the answer to this, because this would take depth recording and this would seriously impair the success of this kind of study. One can only speculate as to whether or not this is so-called secondary, bilateral synchrony, wherein a focal cortical lesion activates the subcortical pacemaker. I remind you that in 28 of our 40 families we have not been able to discover evidence for a familial abnormality. Consequently, in that group of 28 families there may be some patients who have focal structural cortical lesions. I think this certainly must be the kind of mechanism which is operating in the child with the mixed glioma in the left frontaI lobe, whose EEG you saw showing paroxysmal bilateral spike-wave discharges. In this situation one must postulate that the focal hemisphere lesion activated a subcortical pacemaker which produced the diffuse spike-wave discharges which, incidentally, are indistinguishable from pure, if there is such a thing, centrencephalic epilepsy. I think all of these findings are, among other things, a commentary on the limited repertoire of the scaIp encephalogram. DR. EDWARD F. R A ~ , 123 Garden St., Needham 92, Mass. Dr. Bray, I think this is a very thought-provoking presentation. There are two questions which come to mind. One is that some years ago there was described the phenomenon of migration of spike foci in children with age, and this observation was made as the result of the fact that some children with seizure disorder and focal spikes had surgical removal of the area of focal discharge, only with the result subsequently that the spike foci had migrated and the children sometimes persisted in having seizures. As an outgrowth of this, it was noted that children without operation also had migration of spike loci. I wondered if in any of these families that you have studied you had the opportunity to do serial EEG's over the years to find out whether the origin of these spikes that you now found in the temporal lobe began typically in the occipital lobe or in any other area. Second, I wondered if in studying so many persons you came upon any clues as to what may differentiate those persons who have spike loci seizures from those who have spike foci and no clinical manifestations of seizure disorder. DR. BRAY. Let me answer the last question first. We have no idea why some families have seizures and some do not. In a large study by Earl Walker of people who have gunshot wounds of the brain, only 35 per cent of them develop epilepsy. Obviously, the etiology of any seizure is multifactorial, there being a whole host of physiologic, biochemical factors impinging upon the nervous system to vary the state of consciousness. I think it should be said that to my knowledge there is still no convincing evidence about the basic biochemical or molecular defect in this disorder. Now, the question of migratory and shifting
Abstracts
1083
spikes has, of course, been hammered at for a long time. I think Dr. Fred Gibbs was among the first to emphasize this. Dr. Gibbs has postulated that the spike actually moves forward. He takes young infants and finds spikes in the occipital lobes which "migrate forward" on the cortex. This concept has never been very appealing to me. I would rather think that this phenomenon resuits from a slowly developing organ, the brain. The situation is very complex. For example, the neuron population is changing a great deal; the rate of myelination is changing a great deal, and, of possible importance, the temporal lobe is the last to become fully myelinated. This may be one of the reasons why a high percentage of spike discharges from the scalp over the temporal lobe is seen in adults. In order to answer Dr. Rabe's second question, I would like to show this final slide which is a graphic presentation of longitudinal EEG findings in some of these patients. You will note considerable variation in many individuals. At times their recordings will show focal epileptiform discharges. Later these abnormalities will disappear. In some cases they remain absent but in other cases, as you see, the epileptiform abnormalities will reappear again. We feel that observations of this type, based upon longitudinal studies, are important. We have called this "sampling error" which may or may not be the best term.
13. Effect o/prior administration of 7S and 19S antibody on active antibody formation Chester W. Fink and Joseph LoSpalluto, e Departments of Pediatrics, Biochemistry, and Internal Medicine, University of Texas Southwestern Medical School, Dallas, Texas In an earlier report, inhibition of antibody formation by placentaIly transmitted 7S typhoid antibody was demonstrated. In the present study the antibody response to typhoid H antigen has been examined in the rabbit following prior administration of either 7S or 19S antibody. Little or no difference between control animals receiving saline or normal rabbit serum and those receiving 19S antibody was observed. The rabbits receiving 7S antibody, however, showed both a diminished total antibody titer and a marked delay in the appearance of 7S antibody following typhoid H immunization. The differences between groups remained, even when low titers of infused 7S were compared with high titers of infused 19S antibody. Similar differences were found when typhoid H complexes in antibody excess were administered. The results indicate that the 19S antibody exerts little or no inhibitory effect on antibody formation. The specific agglutinating activities of 7S and 19S typhoid H antibody were found to be in the same order of magnitude.
1 0 84
Society for
Pedialric Research
December 1964
In addition, premature infants were immunized with typhoid-paratyphoid vaccine following exchange transfusion. Infants who had no detectable maternally transferred antibody were utilized. In 3 out of 4 infants 19S antibody acquired during exchange did not inhibit active antibody formation, whereas 7S antibody received by exchange completely inhibited it in all of 13 infants. Infants receiving no antibody, either from the mother or by exchange, responded to immunization in a normal fashion. DISCUSSION DR. HORAGE L. HODES, Mt. Sinai Hospital, 11 E. IOOth St., New York 29, N. Y. This is a very interesting paper, describing work which is very beautifully done, and we are indebted again to Dr. Fink for his contributions in this field. I think his suggestion that what we now end up with is the vestigial remains of a less efficient mechanism is a very attractive idea. The 19S antibody-forming is less than optimal in some other ways which we might mention very briefly. One is that the mechanism has a very short memory. In order to keep making 19S antibody, one must continue to receive the antigenic stimulus at short intervals. If the interval is proIonged, formation of 19S antibody stops. It is also less efficient in that 19S antibody does not diffuse so well as smaller antibodies do. I think Dr. Fink's idea that the 19S antibody mechanism is a vestigial thing is a very good one. It is possible that it had something to do with the survival of Homo sapiens as compared with the Pithecanthropus and even the later Neanderthal man, DR. FINK. Hodes.
Thank
you for your comments~
Dr.
DR. E. R. STIEHM, University o[ Cali[ornia Medical Center, San Francisco, Call[. One of the factors important in inhibiting antibody synthesis by administered antibodies would be the duration in which the antibody lasted in the system. From studies on humans, it is felt that the half-life of the 7S antibody is around 30 days and the halflife of the 19S antibody is perhaps 5 days. The half-life of rabbit gamma globulin 7S is about 7 days. I wonder if you have any information on the half-life of your 19S rabbit antibody, and if this might be a possible explanation for the failure of inhibition by the 19S a n t i b o d y - - t h a t it is just not there very long. DR. FINK. We have not done any studies on this ourselves, but there is some information in the literature. It is reported that 7S rabbit gamma globulin has a half-life of about 5 days, whereas the half-life for 19S is about 3 days. We have noted in previous and in the present experiments on humans that infants who have passively transferred 7S antibody may not produce any specific antibody for as long as 6 months after antigen administration. During at least 5 months of this time the passively transferred antibody is below detectable levels. I do not think the shorter halflife of 19S antibody explains our results, although it may be of some significance.
The h u m a n is a better subject for this type of experiment than the rabbit because of the longer half-life of the infused antibody and because the inhibition by 7S is complete rather than partial. However, it was difficult to find infants who had received exclusively 19S antibody during exchange transfusion. The few I have shown were atl that were found in three years of checking all our exchanges. DR. RICHARD T. SMITI-I, Department o/ Pediatrics, University o[ Florida, GainesviUe, Fla. Without in anyway detracting from these data, I must disagree with your interpretation and to some extent with the remarks of Dr. Hodes. Imputing a vestigial role for gamma macroglobulins does not agree with the body of data strongly supporting a major functionaI significance for macroglobulin antibodies in all higher vertebrates and mammals. Their property of preparing bacteria, effete red blood cells, and other particulate matter for phagocytosis is a role in which they appear more efficient than other classes of antibodies. They apparently can be formed when the processes which result in 7~-globulin synthesis are repressed or undeveloped. They appear very early in ontogeny. Dr. John Robbins has recently reported, on the other hand, that macroglobulins do not neutralize diphtheria toxin at all as compared with macroglobulin components directed toward the purified toxin. i n the family of immunoglobulins, each class, in all probability, has special functions and attributes. For example the YA (beta-2A) globulins are the principal secretory form of y-globulin, the 7• (macroglobulins) globulins being limited to the circulation, and the gamma-G globulins (y~) are distributed more widely in the extracellular fluid space. The assumption that macroglobuIins are vestigial and not necessary to the survival of Homo sapiens is really much too speculative to fit the data. DR. HODES. I should like to answer Dr. Smith since I was included in his discussion. The word "vestigial" applied to an organ (or to a function) does not necessarily mean that it is not useful, or that it is on its way out. Anthropologically speaking, I would suggest only that these are different mechanisms, that the 19S is earlier and associated with different phenomena. For example, the appendix is getting smaller and in that sense is vestigial. The h u m a n nose is smaI1 as compared with that of the chimpanzee, but it is still useful. It is in this sense that I used the term "vestigial." DR. FINK. I think there may be other factors that lend credence to this possibility. Phylogenic studies show that the more primitive forms have proIonged 19S production and in some the 19S form of antibody may be predominant. It is also interesting to speculate about the type of antigens which resuIt in predominantly 19S production in humans, an example of this type being typhoid O antigen. These antigens seem to be of the polysaccharide or lipopolysaccharide variety. We have done some preliminary studies in this area.