scholarly journals IMMUNOLOGIC STUDIES OF WATER-SOLUBLE HUMAN AMYLOID FIBRILS

1969 ◽  
Vol 130 (4) ◽  
pp. 797-808 ◽  
Author(s):  
Edward C. Franklin ◽  
Mordechai Pras
Keyword(s):  
Human Serum ◽  
Comparative Studies ◽  
Amyloid Fibrils ◽  
Complement Fixation ◽  
Cross Reactivity ◽  
Water Soluble ◽  
Antigenic Determinants ◽  
Normal Tissues ◽  
Absorption Studies ◽  
Normal Human

Eight preparations of soluble amyloid and degraded amyloid (DAM) were compared immunologically. Unlike amyloid fibrils, six of eight preparations of DAM proved to be relatively strong immunogens. Antisera to DAM reacted weakly or not at all with normal human serum or extracts of normal tissues, but were specifically reactive with amyloid fibrils or DAM. Comparative studies of DAM'S from eight different subjects showed some degree of cross-reactivity among them, yet demonstrated that they were not identical. Similar conclusions were obtained by quantitative precipitin and complement fixation analyses. Comparison of the amyloid fibrils with the homologous DAM by complement fixation and absorption studies demonstrated the existence in DAM of antigenic determinants that were lacking or inaccessible in the native fibrils. A search for amyloid precursors and antibodies to amyloid in the sera of 12 patients proved unsuccessful.

scholarly journals Antibodies to shared idiotypes as agents for analysis and therapy for human B cell tumors

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 430-436 ◽  
Author(s):  
FK Stevenson ◽  
M Wrightham ◽  
MJ Glennie ◽  
DB Jones ◽  
AR Cattan ◽  
...  
Keyword(s):  
B Cell ◽  
Human Serum ◽  
Tumor Cells ◽  
B Lymphocytes ◽  
Cross Reactivity ◽  
Normal Human Serum ◽  
Lymphoma Cells ◽  
Tumor Progress ◽  
Normal Human ◽  
Human B Cell

Abstract Monoclonal anti-idiotypic antibodies generated against idiotypic immunoglobulin (Ig) of neoplastic B lymphocytes can be selected from growing hybridoma clones by their ability to recognize idiotypic but not normal IgM. This group of antibodies can be subdivided into those that bind to the target tumor cells in the presence of normal human serum (approximately 85% of the clones) and those in which binding is inhibited by serum (approximately 15%). The former appear to be specific for private idiotypic determinants whereas the latter recognize cross-reacting idiotypic determinants. Such cross-reactivity is reflected both in recognition of a small percentage of normal Ig and also in binding to other lymphomas. The anti-idiotypes specific for private determinants can be used for therapy, with only idiotypic Ig secreted by tumor cells able to block its access to cells. The cross- reacting anti-idiotypes will face in addition the barrier of the proportion of normal Ig with which it reacts. The attraction of using a single monoclonal reagent for more than one patient has led us to develop an assay that measures the level of such blocking and to propose that those recognizing less than 30 micrograms/mL of normal Ig could be placed in a panel for possible therapy for several patients; less restriction need apply to antibodies for monitoring tumor progress. The assay is described, and examples of such antibodies raised against lymphoma cells from two patients are given together with comparisons with them of anti-idiotypes specific for private determinants.

scholarly journals INTERRELATIONSHIP BETWEEN THE Rh SYSTEM AND THE A B SYSTEM

Blood ◽  
1948 ◽  
Vol 3 (Special_Issue_Number_2) ◽  
pp. 66-79 ◽  
Author(s):  
ERNEST WITEBSKY ◽  
MRS. LIVIA BLUM ◽  
MISS DORIS HOWLES ◽  
MISS HELEN WARD
Keyword(s):  
Human Serum ◽  
Blood Group ◽  
Saline Solution ◽  
High Titer ◽  
Water Soluble ◽  
Normal Human Serum ◽  
Experimental Conditions ◽  
Blocking Antibody ◽  
Group A ◽  
Normal Human

Abstract The isoantibodies anti-A and anti-B which are described differ in several respects from those occurring in normal human serum. This type of antibody has first been observed in the serum of an Rh negative woman who exhibited a history of erythroblastosis. Her husband belonged to the subtype Rh1 and to the blood group A. The patient’s serum completely neutralized with A and B substances still agglutinated strongly the husband’s cells provided normal human serum replaced physiological saline solution as a diluent for all dilutions. The impression was thus created that an Rh blocking antibody was responsible for the agglutination observed. It could be shown, however, that the abnormal antibody present in this patient’s serum was not an Rh antibody at all but instead, an antibody directed against the A property. This type of anti-A antibody resembles the Rh blocking antibody in many respects. It becomes manifest only if undiluted human serum is used as a diluent. Surprisingly enough this antibody agglutinated cells of group A, although the amount of AB substances added to the serum was sufficient to neutralize completely the isoagglutinin anti-A under normal conditions in which saline solution is used as a diluent. This anti-A antibody therefore cannot be neutralized as easily as the normal isoagglutinin anti-A. For its neutralization much larger amounts of the blood group specific substances are apparently necessary. The patient’s serum fixed complement when mixed with material containing water soluble A substance, in contrast to the normal isoantibody anti-A which failed to do so. The titer of isoantibodies found in the patient’s serum upon titration in saline solution was not extensively high and, as a matter of fact, was average. It is therefore felt that an extremely high titer is neither a necessary requirement nor proof of isoimmunization toward the A and B factors. Another interesting characteristic of the peculiar anti-A antibody occurring in our patient’s serum was the fact that it was essentially an anti-A1 antibody. The difference in agglutination between A1 and A2 cells respectively becomes manifest if normal serum is used as a diluent instead of saline solution. This difference becomes even more marked after neutralization of the patient’s serum with A and B substances. During the course of Mrs. Bong’s pregnancy the special anti-A antibody described did not increase but rather decreased in strength. However, even after delivery the antibody was demonstrable for at least several weeks although we had no opportunity to examine the patient’s serum further. That one must be very careful in contributing any pathological significance to isoantibodies anti-A or anti-B, even of the type described, is evident from the fact that the patient was delivered of a perfectly normal baby belonging to the blood group O and being Rh negative. Whether the difficulties experienced by the patient in previous pregnancies were due to sensitization toward the Rh factor or to the A factor cannot be decided. Antibodies anti-A and anti-B of the type reported were also found in the sera of patients who had received large amounts of pooled plasma or O blood conditioned with A and B specific substances. Again the anti-A antibody occurring in the serum of these patients was mainly directed against the A1 property. Under the experimental conditions described in this paper, such a serum can be used for the differential diagnosis of the subgroups A1 and A2 and constitutes a sensitive reagent for the recognition of the differences occurring within the A factor. With the aid of such a serum only 10 per cent of A cells were found to belong to subgroup A2, 75 per cent to A1, and 15 per cent were considered to be of the intermediate type. No subgroups were found so far in human cells of group B.

scholarly journals THE ANTIGENIC AND MOLECULAR ALTERATIONS OF C3 IN THE FLUID PHASE DURING AN IMMUNE REACTION IN NORMAL HUMAN SERUM

1971 ◽  
Vol 134 (3) ◽  
pp. 656-680 ◽  
Author(s):  
R. E. Spitzer ◽  
A. E. Stitzel ◽  
V. L. Pauling ◽  
N. C. Davis ◽  
C. D. West
Keyword(s):  
Human Serum ◽  
Immune Complex ◽  
Fluid Phase ◽  
Polyacrylamide Gel ◽  
Normal Human Serum ◽  
Antigenic Determinants ◽  
Complement Components ◽  
Molecular Alterations ◽  
Normal Human ◽  
Kinetics Of

During the reaction of an immune precipitate with fresh human serum, C3 undergoes a number of molecular alterations with the formation of conversion products differing from those obtained when purified components react. Those products which remain in the fluid phase, the subject of the present paper, have been identified by their reaction with monospecific antisera to the three antigenic determinants of C3, A, B, and D, after electrophoresis in agar or polyacrylamide gel. When purified C3 reacts with EAC1,4,2, C3i is found in the fluid phase. C3i, a loose complex of C3a and C3b, is in a conformational state whereby only the A and D antigens, present on its C3b portion, will consume antibody. The B antigen, present on the C3a portion of C3i, is unavailable for combination with antibody until C3i dissociates. In the fluid phase of the reaction of an immune precipitate with whole serum, C3i, C3a, and C3b, formed when purified components react, cannot be found. Instead the end products of the reaction appear to be C3c, which contains the A antigen, and C3d, which contains the D antigen. C3c and C3d are similar to the ß1A and α2D produced by the aging of serum but differ in their mobilities in acrylamide gel and in agar. The C3c and C3d generated by an immune precipitate also differ slightly from the C3c and C3d produced by the reaction of trypsin with C3 in whole human serum. As human serum reacts with an immune complex, native C3 appears to undergo a primary alteration before conversion. This alteration results in a molecular species of C3 which is labile at 56°C for 30 min, fails to expose additional A and D antigenic sites upon aging, and which forms ß1A and C3d rather than ß1A and α2D during aging. In addition to this altered form of native C3, a new conversion product, C3x, is formed as whole serum reacts with an immune complex. C3x is not found in systems utilizing pure complement components. C3x is like C3 in that it bears all three antigenic determinants but differs in that it has a slightly faster mobility in polyacrylamide gel than does native C3. C3x is not only found in the fluid phase but is also bound to the immune precipitate. Finally, the fluid-phase kinetics of each of the antigens of C3 have been determined as normal human serum reacts with an immune precipitate. These illustrate that nearly the entire population of native C3 molecules undergoes conversion rapidly as manifested by the disappearance of the B antigen from the fluid phase. Moreover, the kinetics of the fluid-phase A and D antigens reflect that the conversion of C3 in serum is quantitatively not the same as when purified C3 reacts with C4,2.

scholarly journals IMMUNOCHEMICAL STUDIES ON BLOOD GROUPS

1971 ◽  
Vol 134 (2) ◽  
pp. 439-457 ◽  
Author(s):  
Carlos Moreno ◽  
Arne Lundblad ◽  
Elvin A. Kabat
Keyword(s):  
Complement Fixation ◽  
Blood Groups ◽  
Structural Basis ◽  
Antigenic Determinants ◽  
Absorption Studies ◽  
Inhibition Studies

The immunochemical properties of purified A1 and A2 glycoproteins have been compared to ascertain whether their antigenic determinants differ. Quantitative precipitin and complement-fixation studies using several anti-A sera as well as purified γG anti-A antibodies clearly showed a specificity difference. This was also supported by absorption studies: A2 substance specifically removed antibodies reacting with A2 substance leaving anti-A1 activity. A1 substance was more effective than A2 substance in dissolving an A1 anti-A1-specific precipitate. Purified γM anti-A hemolyzed A1 cells more readily than A2 cells. Inhibition studies using mono- and difucosyl type 2 A-active oligosaccharides showed that type 2 difucosyl receptors are present in A2 substance. The structural basis for the specificity difference between A1 and A2 would appear to be that A2 substances lack type 1 A determinants; this would account for the observed higher H and Leb activity in A2 substances.

scholarly journals A Study of Non-specific Complement-fixation with particular reference to the Interaction of Normal Serum and certain Non-antigenic substances

1928 ◽  
Vol 28 (2) ◽  
pp. 172-197 ◽  
Author(s):  
T. J. Mackie ◽  
M. H. Finkelstein
Keyword(s):  
Human Serum ◽  
Complement Fixation Test ◽  
Complement Fixation ◽  
Normal Serum ◽  
Fixation Test ◽  
Normal Human Serum ◽  
Rabbit Serum ◽  
Considerable Proportion ◽  
Normal Human ◽  
Fixation Reaction

1. When a solution of commercial peptone is substituted for antigen in a complement-fixation test with the unheated normal serum of certain species (man, ox, sheep, horse, rabbit, white rat), a definite fixation reaction occurs both at 37° C. and at 0° C. In the ox, sheep, horse and rabbit this property of serum is partially stable at 55° C., but normal human serum and the serum of the white rat are inactive after heating at this temperature. The property is resident mainly in the carbonic-acid-insoluble globulins of the serum.2. The same results are obtained when ethyl alcohol diluted with several volumes of normal saline solution is substituted for antigen in a complement-fixation test with normal serum.3. Analysis of these reactions shows a close correspondence with complement-fixation by the sera of normal animals plus the Wassermann “antigen”—the Wassermann reaction of normal animals.4. Marked complement-fixation effects are also obtained with heated normal serum of the rabbit, ox, sheep, horse plus cholesterol suspension, and particularly cholesterolised-peptone, these effects occurring in parallel with those produced by serum plus alcohol-saline, peptone solutions and the Wassermann “antigen.” The heated normal serum of the pig, white rat and guinea-pig do not exhibit these reactions, and the same applies to heated normal human serum. Unheated pig serum fails to react. Such results also elicit a close relationship between these non-specific reactions and the Wassermann reactions of normal animals.5. The reacting property is absent from the serum (heated and unheated) of young rabbits during the first 2 to 3 weeks of life, but appears soon after this (e.g. by the 37th day) and is progressive in development. Its development in early life runs parallel to that of the natural haemolytic property of the serum for sheep's blood (due to a natural antibody-like substance). The two properties are, however, independent as illustrated by absorption tests.6. Besides the agents referred to above as capable of fixing complement along with normal sera, other substances possess a similar property, e.g. certain alcohols, sodium oleate, tissue proteins, certain amino-acids and sodium nucleate. Commercial peptone purified by precipitation with alcohol is equally active with the original material. Cholesterolisation of these agents may yield a product whose activity is greater than that due to summation of effects.7. Wassermann-positive and -negative human sera have been tested in the complement-fixation reaction with certain of these “pseudo-antigens,” viz. alcohol-saline, peptone, cholesterol, and cholesterolised-peptone, but a uniform parallelism has not been demonstrated between the reactions with these agents and the Wassermann effect. Some Wassermann-positive sera react also with alcohol-saline, peptone, cholesterol and cholesterolised-peptone, while sera from selected normal persons are quite inactive. A considerable proportion of Wassermann-positive sera yields definite complement-fixation with cholesterol and cholesterolised-peptone; a small proportion of Wassermann-negative sera reacts with these agents.8. The thermolability of the serum principles acting with various “pseudoantigens” has been studied by testing unheated serum and serum heated at temperatures ranging from 46° to 60° C. Two types of thermolability curve have been demonstrated with different specimens of rabbit serum: (1) a more or less progressive weakening of the various reactions with inactivation at 60° C.; (2) inactivation of the effects with Wassermann “antigen,” alcoholsaline and cholesterol at 50–52° C., activation of the effects with the Wassermann “antigen” and cholesterol at 54–56°C. and inactivation again above 60° C.; in this case the curves for peptone and cholesterolised-peptone do not show such double inactivation. Unheated normal human serum yields reactions with the various agents (including the Wassermann “antigen”) but inactivation occurs at 50° to 54° C. whereas certain syphilitic sera yield thermolability curves somewhat similar to type (1) of rabbit serum, with inactivation at 60° C. or over.

scholarly journals Antibodies to shared idiotypes as agents for analysis and therapy for human B cell tumors

Blood ◽  
1986 ◽  
Vol 68 (2) ◽  
pp. 430-436
Author(s):  
FK Stevenson ◽  
M Wrightham ◽  
MJ Glennie ◽  
DB Jones ◽  
AR Cattan ◽  
...  
Keyword(s):  
B Cell ◽  
Human Serum ◽  
Tumor Cells ◽  
B Lymphocytes ◽  
Cross Reactivity ◽  
Normal Human Serum ◽  
Lymphoma Cells ◽  
Tumor Progress ◽  
Normal Human ◽  
Human B Cell

Monoclonal anti-idiotypic antibodies generated against idiotypic immunoglobulin (Ig) of neoplastic B lymphocytes can be selected from growing hybridoma clones by their ability to recognize idiotypic but not normal IgM. This group of antibodies can be subdivided into those that bind to the target tumor cells in the presence of normal human serum (approximately 85% of the clones) and those in which binding is inhibited by serum (approximately 15%). The former appear to be specific for private idiotypic determinants whereas the latter recognize cross-reacting idiotypic determinants. Such cross-reactivity is reflected both in recognition of a small percentage of normal Ig and also in binding to other lymphomas. The anti-idiotypes specific for private determinants can be used for therapy, with only idiotypic Ig secreted by tumor cells able to block its access to cells. The cross- reacting anti-idiotypes will face in addition the barrier of the proportion of normal Ig with which it reacts. The attraction of using a single monoclonal reagent for more than one patient has led us to develop an assay that measures the level of such blocking and to propose that those recognizing less than 30 micrograms/mL of normal Ig could be placed in a panel for possible therapy for several patients; less restriction need apply to antibodies for monitoring tumor progress. The assay is described, and examples of such antibodies raised against lymphoma cells from two patients are given together with comparisons with them of anti-idiotypes specific for private determinants.

The C3b Component of Complement Inhibits NK Cell Activation Induced by Rituximab-Coated Target Cells.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2505-2505 ◽  
Author(s):  
Siao-Yi Wang ◽  
Emilian Racila ◽  
Ronald P. Taylor ◽  
George J. Weiner
Keyword(s):  
Human Serum ◽  
Nk Cells ◽  
Cell Activation ◽  
Complement Fixation ◽  
Nk Cell ◽  
Normal Human Serum ◽  
Heat Inactivation ◽  
Target Cells ◽  
Normal Human ◽  
The Relationship

Abstract Rituximab (RTX) is an accepted therapy for B cell malignancies, but there is still much to learn about the mechanisms responsible for the observed responses and the potential interactions between various mechanisms of action. Some studies suggest that complement fixation followed by lysis through the membrane attack complex contributes to the anti-tumor effects of RTX. Other investigations indicate that antibody dependent cellular cytotoxicity (ADCC) mediated by NK cells is central to the response of therapy. In prior studies, we found that RTX-coated target cells activate NK cells as indicated by NK cell modulation of CD16, upregulation of CD54 and CD69, and production of IFNγ. NK activation induced by RTX-coated target cells was dependent on the affinity of multivalent interactions between Fc γ receptors III (CD16) of the NK cell and Fc regions of cell-bound RTX molecules. We used these in vitro assays to assess the relationship between complement fixation, and the ability of RTX-coated target cells to activate NK cells. Normal human serum inhibited the modulation of NK cell CD16, and also blocked upregulation of CD54, induced by RTX-coated target cells. The ability of serum to inhibit NK activation was dose dependent and was abrogated upon heat inactivation. Serum depleted of C1q or C3 also failed to inhibit NK cell activation. The inhibitory activity of serum depleted of these complement components was restored when purified C1q or C3 were added back respectively. In addition, the level of NK cell inhibition was dependent on the amount of C3b deposited on the target cells. An antibody that stabilizes C3b on the target cell surface (3E7, DiLillo et al., Molec Immunol 2006) further enhanced the inhibition of NK cell activation induced by RTX-coated target cells. One possible explanation for these findings is that complement-mediated lysis destroyed the RTX-coated target cells before they had the opportunity to induce activation of the NK cells. To assess this possibility, lymphoma cells were killed, fixed with formaldehyde, and washed prior to their use as target cells. These RTX-coated and fixed target cells were able to induce modulation of CD16 on the NK cells, which was again inhibited by normal human serum. These findings indicate that the observed inhibition of NK activation by complement is unlikely to be a consequence of complement mediated lysis of the target cells. Instead, these data suggest that C3b deposition on RTX-coated target cells inhibits the interaction between the Fc portions of RTX and CD16 on the NK cells, and so limits the ability of RTX-coated target cells to induce NK activation. These results could have implications in our understanding of the relationship between complement fixation and ADCC, and their relative roles in potentiating destruction of malignant cells in the blood and tissues.

scholarly journals COMPARATIVE STUDIES ON THE PHYSICAL CHARACTERISTICS OF THE HERITABLE HAPTOGLOBIN GROUPS OF HUMAN SERUM

1959 ◽  
Vol 109 (1) ◽  
pp. 55-68 ◽  
Author(s):  
A. G. Bearn ◽  
E. C. Franklin
Keyword(s):  
Human Serum ◽  
Comparative Studies ◽  
Normal Human Serum ◽  
Serum Haptoglobin ◽  
Zone Electrophoresis ◽  
Before And After ◽  
Group Variation ◽  
Normal Human ◽  
Group 2 ◽  
Group 1

Ultracentrifugal studies of the α2-proteins of normal human serum before and after the addition of hemoglobin have revealed three separate and clearly distinguishable patterns based on the three major serum haptoglobin groups. Isolation of the three haptoglobin hemoglobin complexes disclosed characteristic patterns for each group. The heterozygote was found to possess electrophoretic and ultracentrifugal components not seen in either homozygote. Separation of normal human serum, to which hemoglobin had been added, by conventional starch zone electrophoresis resulted in three characteristic patterns which also permitted the ready identification of the serum haptoglobin group. The haptoglobin hemoglobin complex from Group 1-1 had the fastest mobility and that of Group 2-2 the slowest. The heterozygote Group 2-1 had an intermediate mobility. Immunologic differences between the haptoglobin hemoglobin complexes of the three groups could not be detected. Some of the variations in the reported electrophoretic and ultracentrifugal patterns of normal human serum could be reasonably ascribed to the group variation of the haptoglobins in man. The physical studies imply that the proteins characteristic of the heterozygote differ in size and charge from those present in either of the two homozygotes and indicate that the genetic control of the synthesis of the serum haptoglobins is probably exceedingly complex.

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