bluuebunny/crossref_metadata_2025 huggingface dataset:
Studies on Synthetic Polypeptide Antigens
Authors: Simonian S. J. , Gill T. J. , Gershoff S. N.
Published: December 2022

Immunization of random bred Charles River CD strain (CR) rats with poly Glu52Lys33Tyr15 (no. 3) in Freund's complete adjuvant resulted in a highly variable antibody response which was most likely due to random genetic fluctuations in the population. Immunization of six inbred (BN, BUF, F344, WF, LEW and ACI) and one systematically random bred (WIST) strain of rats with a variety of synthetic polypeptide antigens resulted in a reproducible antibody response in all strains, but with marked interstrain differences. Studies with (ACI × F344) F1 hybrid rats showed that the ability to make antibody was transmitted as a complex genetic control mechanism which probably involved several genes.

The presence of tyrosine in the antigen was not necessary in order to elicit an antibody response, and its effect on the magnitude of the antibody response was variable and strain-dependent. All strains responded to a D-amino acid polypeptide and showed interstrain differences. Aggregation of an L-polypeptide with methylated bovine serum albumin elicited an increased antibody response in those strains which produced a low titer to the polypeptide alone and reduced interstrain differences in the response. A D-polypeptide aggregate elicited more antibody in all strains except WIST and abolished the interstrain differences in the antibody response.

The antibody response to a spatially ordered, dinitrophenylene (DPE) cross-linked polypeptide was higher, showed increased interstrain differences and showed a different specificity than the antibody response to the parent, linear polypeptide or the (Lys-DPE) and DNP conjugates, which are partial structural analogues of the DPE cross-link. Thus, the DPE cross-linked polypeptide had unique antigenic sites which depended upon the spatial organization of the molecule. In contrast, the presence of amide cross-links in the molecule introduced a relatively small change in immunogenicity and antigenic specificity.

The studies reported here provide evidence that the effects of the various structural aspects of an antigen on immunogenicity and antigenic specificity are under genetic control. The ability to respond to a particular facet of antigenic structure probably involves several genes.
Suckling Mouse Cataract Agent (SMCA) in Mice
Authors: Clark H Fred , Karzon David T.
Published: December 2022

SMCA virus induces cataracts in a high percentage of infected suckling mice. Maximal SMCA-induced cataract rates were obtained only when virus doses of 104.0 ELD50 or greater were inoculated intracerebrally into suckling mice 4 days of age or younger. Virus inoculated by the intravenous or intraperitoneal routes did not produce cataracts. The SMCA-induced cataract rate was mouse strain-dependent. Cataract rates, based on eye examination at age 30 days, varied from 85% in strain C57B1/6Ha mice to 3% in strain A mice. After 30 days, rates decreased in most mouse strains, but in strain A mice the cataract rate gradually increased to a rate of 44% at 1 year (late cataract strain).

Comparative virus assays were performed of the brains, liver-spleen tissues and eyes of SMCA-inoculated strain CFW(A) (30-day cataract rate 46%) and strain A (30-day cataract rate 3%) mice. In each mouse strain virus reached peak titers in all tissues studied in 7 to 15 days. Virus titers then slowly decreased, disappearing from liver-spleen tissues and most eyes by 60 days, but persisting indefinitely (up to 827 days) in the brain. Eye infection rates were similar (88% in A mice, 91% in CFW(A) mice) in both strains of mice. However, in the late cataract strain A mice, virus clearing from eyes and viscera occurred earlier, and peak titers of virus in eyes were lower than in CFW(A) mice.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Klemperer M. R.
Published: December 2022

The antigenicity of human C′2, purified by the method of Lepow, was enhanced by treatment with neuraminidase. A precipitating rabbit antibody to human C′2 was obtained. The serums of four homozygous C′2-deficient individuals from two kindreds failed to form bands of precipitation with this antibody. The serums of 15 heterozygous C′2-deficient from these kindreds contained 30 to 60% of normal C′2 as determined by a quantitative immunodiffusion assay. The normal serum level of C′2 was estimated to be 1.0 ± 0.2 mg%. The sera of heterozygous C′2-deficient individuals from both kindreds gave lines of complete fusion with the serum from a normal individual and with each other, an indication that their protein was not deficient in antigenic determinants. Upon immunoelectrophoretic analysis, the precipitation band formed with C′2 in normal serum had the mobility of a β1 globulin. Serums obtained from indivduals with attacks of hereditary angioneurotic edema form the C′2 precipitation band in a more cathodal position than normal when subjected to immunoelectrophoresis.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Shin H. S. , Pickering R. J. , Mayer M. M. , Cook C. T.
Published: December 2022

C′5 was isolated by isoelectric precipitation, Pevikon electrophoresis and chromatography on DEAE cellulose and hydroxyapatite. The sedimentation coefficient was 7.9 S for purified C′5 and for C′5 in serum. Molecular weight of 200,000 was estimated on Sephadex G 200. On reaction with EAC′4,2a,3, some C′5 was fixed (ca. 1/10), and the remainder was inactivated. In accord with this, at low ionic strength, 10 molecules were required for lysing one erythrocyte. At high ionic strength, the reaction rate was lower. The efficiency of site formation was also lower, due to dissociation of cell-bound C′5. The dissociated C′5 either recombines with EAC′4,2a,3 or becomes inactivated, the former (site-to-site transfer of activated C′5) being favored at high SAC′4,2a,3 concentration. There are two mechanisms of C′5 dissociation. One, spontaneous dissociation, is favored at high ionic strength and occurs even at 0°C. The other, a consequence of decay-release of C′2a, is favored by elevated temperature and occurs even at low ionic strength.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Opferkuch W. , Ringelmann R. , Klein P. G.
Published: December 2022

When guinea pig euglobulin is exposed at 37°C to a pH of 7.8 and an ionic strength of µ = 0.15, an anticomplementary factor is generated. This factor can be purified. The respective fraction rapidly inactivates purified guinea pig C′4 and hydrolyzes TAMe; it is not active against purified guinea pig C′2, neither alone nor in the presence of C′4. The characteristics of the C′4 inactivation reaction are the following: The reaction velocity decreases markedly and reaches a minimum after 15 min. The rate of C′4 inactivation is not markedly increased by higher temperatures. The C′4-inactivating capacity of the euglobulin fraction is consumed during its interaction with C′4. On the basis of these traits, a titration of the inactivator can be performed. The kinetics of the process of generation of the C′4 inactivator are studied. The data do not indicate that C′1 acts as a precursor for the C′4 inactivator.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Ruddy S. , Austen K. F.
Published: December 2022

C′3 Inactivator (C′3 INA) purified from human serum has been measured by its ability to destroy the activity of C′3 bound to the cellular intermediate EAC′143. Dilutions of C′3 INA are incubated at 30°C for 30 min with cells bearing limited numbers of C′3 sites. The residual cell-bound C′3 is then detected in a stoichiometric hemolytic assay: the cells are treated with excesses of functionally pure C′2,5,6 and 7 for 30 min at 30°C which converts the remaining active SAC′143 to the state SAC′1423567, and these are in turn converted to S* by treatment with excesses of C′8 and C′9 for 60 min at 37°C. The proportion of C′3 sites inactivated can be calculated and shown to be a function of C′3INA input. The activity of C′3 INA is not influenced by the presence of C′1 or C′2 on the cell in that it is equally effective against SAC′1423, SAC′143 or SAC′43.
Abstracts of Papers Presented at the Third International Complement Workshop, Harvard Medical School, Boston, Massachusetts, June 3–5, 1968
Authors: Jensen J. A. , Sigel M. M. , Ross G. D.
Published: December 2022

Sensitization of sheep E with nurse shark (N) serum resulted in the formation of EAN if the serum was heat-inactivated or reacted in 0.04 M EDTA at 30°C, of EANC′1N if the serum was freeze-inactivated or reacted in 0.04 M EDTA at 0°C. Guinea pig (g.p.) complement or sup. I readily lysed EANC′1N but not EAN due to a complete C′1 incompatibility, which could also be demonstrated in vivo: in the absence of C′1N, the then non-complement-fixing antibody was not only tolerated by guinea pigs, but it protected them against lethal Forssman shock. Shark C′1 lysed EARa and its intermediate complexes with g.p.-C′ only to the extent of its contamination with antibody; a noteworthy exception was the complex EARaC′1–C′8 g.p. A large variety of Forssman positive and negative erythrocytes were agglutinated and sensitized by the shark's natural antibodies. Young animals had much less antibody but a fully reactive complement system. A powerful, nontoxic inactivator of mammalian C′4 could be isolated and separated from antibody, C′1, and most other serum proteins by gel filtration of low ionic strength precipitates from normal shark serum.
Unequal Association of Mouse Allotypes with Antibodies of Different Specificities
Authors: Weiler Eberhardt , Weiler Ivan Jeanne
Published: December 2022

The experiments were concerned with whether in F1 heterozygous mice the two parental allotypes are expressed in constant proportions among antibody-producing cells or whether their proportions vary from mouse to mouse.

Poly-L-alanine and poly-d-alanine were used as non-crossreacting haptenes. Antibody production by individual spleen cells was recorded in a plaque assay procedure for which the poly-alanine haptenes were linked to rabbit antibodies (IgG) against sheep red blood cells, which in turn were attached to sheep red blood cells. The allotypes of antibodies produced by the individual spleen cells were determined by the facilitation reaction employing reciprocal allotype antisera prepared in the parental mouse strains.

A wide fluctuation was found from mouse to mouse in the proportions of cells producing antibodies of the two parental allotypes, especially when the “weaker” haptene, poly-L-alanine, was used as the antigenic determinant. The distribution of the allotype ratios as recorded for poly-L-alanine was heterogeneous in the group of mice studied. In mice injected with both poly-D- and poly-L-alanine, the allotype ratios recorded with poly-L-alanine were not correlated with the allotype ratios recorded with poly-D-alanine.

The results are interpreted to mean that the cellular determination to produce antibodies of a given specificity, and the decision about which of two allelic chromosome regions will be active, are associated and are transmitted together in cell lines or clones.
Communications
Authors: Torisu Motomichi , Sonozaki Hidekichi , Ito Koji , Shiraishi Satoshi
Published: December 2022

It is not yet clear whether or not skin-sensitizing antibody requires the complement system when it fixes to the cutaneous site in the Prausnitz-Küstner (PK) reaction. Attempts to analyze in vivo the role of complement in various immunologic phenomena have been carried out employing specific inhibitors of complement components (1). The effect of C′3 inactivator and Cu-chlorophyllin (CuC) on the PK reaction was examined. The results are described in this report.

C′3 inactivator is the substance which inactivates the C′3 site of EAC′1423 both in immune adherence and in immune hemolysis (2). It was purified from human serum using column chromatography on diethylaminoethyl (DEAE)-cellulose and carboxymethyl (CM)-cellulose, followed by Sepadex G 200 and Sephadex G 200 super-fine gel filtration (3). The titer of C′3 inactivator was measured by a method previously described (4). Sodium copper chlorophyllin was shown to be a potent inhibitor in both immune hemolysis and passive cutaneous anaphylaxis (PCA) (1, 5).
Comparative Sensitivities of the Hemolytic Plaque-in-Agar and Microchamber Assays for Detection of Rabbit Immune Cells
Authors: Merchant Bruce , Petersen Bruce
Published: December 2022

The microchamber and the plaque-in-agar localized hemolysis techniques were used simultaneously to assay rabbit immune spleen cell suspensions for high hemolytic efficiency plaque-forming cell activity. Direct plaque-forming cell activities both for erythrocyte and for para-azo-benzenearsonic acid determinants were closely comparable by both methods, but plaque size frequency distribution data revealed the microchamber procedure to be potentially the more sensitive. Plaque size distribution data also indicated that for those cells recovered in suspensions the assay conditions permitted detection of the complete population of high hemolytic efficiency plaque-forming cells by either the microchamber or the plaque-in-agar techniques. Accordingly, for relevant plaque-forming cells recovered in suspension, relative assay efficiency closely approached absolute assay efficiency.
The Effect of Passive Antibody on Protein Synthesis in Lymph Node Cells of Immunized Rats
Authors: Greenberg Leonard J. , Uhr Jonathan W.
Published: December 2022

The incorporation of leucine into popliteal lymph node cells was studied in immunized rats some of which received passive antibody 2 days after antigen. In rats receiving antigen alone, there was a phase of increased incorporation into the cytoplasm beginning on day 4, which corresponded to the phase of rapid antibody synthesis. The administration of passive antibody prevented this phase but transiently stimulated increased incorporation of leucine, which reached a peak on day 4. It was suggested that this transient phase represents stimulation of lymphocytes by antigen-antibody complexes.
Production and Properties of Mouse Antihapten Antibodies
Authors: Mattioli Carlos A. , Yagi Yasuo , Pressman David
Published: December 2022

Mice of inbred strains, despite their genetic homogeneity, produced antibodies with several types of molecular heterogeneity. A simple hapten, the p-azobenzoate group, elicited antibodies of the three γG classes. Furthermore, individual mice of the same strain produced anti-p-azobenzoate antibodies of widely different affinity.

The antibodies directed against p-azobenzoate groups of a conjugate had a faster electrophoretic mobility than did the antibodies directed against the carrier protein of the same conjugate. This striking characteristic was found in all the strains tested and it was independent of the immunoglobulin class or carrier used.

For the production of antibodies against the p-azobenzoate hapten, hemocyanin seems to be a better carrier protein than bovine serum albumin. Peritoneal fluid was used as the source of antibodies and myeloma globulins. Abundant amounts of peritoneal fluid could be almost always induced by the intraperitoneal inoculation of Ehrlich ascites tumor cells, whereas the inoculation of Freund's adjuvant failed to produce ascites in some strains. Antibodies were recovered from ascitic fluid using insoluble specific immunoadsorbents.
Localization of 125I-Labeled Antigen in Germinal Centers of Mouse Spleen: Histologic and Ultrastructural Autoradiographic Studies of the Secondary Immune Reaction
Authors: Hanna M. G. , Szakal A. K.
Published: December 2022

Histologic and ultrastructural studies were carried out on the process of the localization of antigen in splenic germinal centers during the secondary immune reaction in mice. The magnitude and regularity of the developed plasma membrane infoldings of antigen-retaining reticular cells in germinal centers are described. The key finding was that these specifically developed plasma membrane infoldings are degraded during the early intervals of the secondary response. This response occurred in conjunction with a loss of label and an apparent loss of function of the existing antigen-trapping mechanism in splenic germinal centers. We suggest that the described cytotoxic sequelae could be a result of complement fixation and that such a regulatory mechanism would be essential to prevent further contact of immunologically competent cells with antigen, while clearing the centers for their development against other antigens. A conformity of vesicles was observed in the cytoplasm of the parenchymal cells of the germinal centers and in the plasma membrane infoldings of the antigen-retaining reticular cells.
Effects of Radiation on Direct and Developed Spleen Hemolysin Plaque-Forming Cells of Mice
Authors: Uyeki Edwin M. , Klassen Robert S.
Published: December 2022

The effects of 500 r whole body γ radiation on direct and developed hemolysin plaque-forming cells in spleen tissue and on serum hemolysin levels of LAF1 mice were investigated. The major findings were: Fourteen-day assay involving a single antigen injection: When radiation was administered 2 days before or 3 days after antigen, spleen hemolysin plaque-forming cell production was markedly lowered when compared with spleen hemolysin plaque-forming cells of non-radiated animals. With this injection scheme, the principal cell type was the developed hemolysin plaque-forming cell. Spleen cellularity of radiated animals was significantly below that of non-radiated control animals.Fourteen-day assay involving two antigen injections: A second antigen injection 6 days after radiation stimulated antibody-forming cell production so that the total spleen hemolysin plaque-forming cells of the once-radiated animals were comparable to non-radiated control animals. Radiation, administered after the second antigen, markedly inhibited hemolysin plaque-forming cell production as well as spleen cellularity.Twenty-eight-day observation period involving multiple antigen injections: The effects of a single dose of 500 r administered to LAF1 mice 3 days after an initial antigen injection and given a total of two, three and four antigen injections at weekly intervals and analyzed at 14, 21 and 28 days, respectively, showed no difference from non-radiated animals in their ability to respond to antigenic stimulation. In these animals, spleen cellularity was consistently lower than in non-radiated animals.
Effects of Chronic Bronchopulmonary Disease and Aging on Human Nasal Secretion IgA Concentrations
Authors: Alford Robert H.
Published: December 2022

Normal subjects and hospitalized chronically ill patients who did or did not have bronchopulmonary disease were compared according to nasal wash protein and immunoglobulin concentrations. Persons with chronic bronchopulmonary disease were not different from other chronically ill patients in these respects. However, both groups of patients differed statistically from normal subjects in nasal wash IgA concentrations. This difference was accounted for on the basis of decreasing nasal secretion IgA concentration with advancing age. This decrease was considered to represent one of several alterations in immunologic competence associated with aging.
Immunoglobulin Production in Bronchial Mucosa and Bronchial Lymph Nodes, Particularly in Cystic Fibrosis of the Pancreas
Authors: Martinez-Tello Francisco J. , Braun Dietmar G. , Blanc William A.
Published: December 2022

The three main serum immunoglobulins were demonstrated immunohistologically in plasmacytes of the human bronchial mucosa. The majority of the Ig-producing cells produce IgA and only a minority of cells produce IgM. There was no immunoglobulin production in three newborn infants and one agammaglobulinemic child.

The number of plasma cells containing immunoglobulins increases significantly in the bronchial mucosa in chronic bronchopulmonary disease. Those containing IgA are the most abundant followed by those containing IgG. Cells containing IgM are not increased in number.

Parallel immunohistochemical studies of bronchial lymph nodes have shown an increased number of Ig-containing cells in chronic inflammation. In contrast to bronchial mucosa, IgG-containing cells are by far the most abundant.

Immunodiffusion studies suggested the presence of 11 S IgA in bronchial washings. Using immunofluorescent staining the “secretory piece” was found in the bronchial epithelium and bronchial glands as well as in bronchial secretions. The secretory piece was demonstrated immuno-histologically in the bronchial epithelium, glands and secretions of newborn infants and of an agammaglobulinemic patient.

In the particular case of CFP, immunologic and immunohistochemical studies suggest that there is no defect of the production and secretion of IgA in the bronchial mucosa. On the contrary, there is high IgA production. The role of IgA in the local defense mechanism in chronic bronchopulmonary processes is discussed.
Antibody-Producing Cells in Rabbits Injected with Soluble BSA
Authors: Daniels J. C. , Weigle William O.
Published: December 2022

A hemolytic plaque technique has been modified for detecting antibody-producing cells in lymphoid tissues of rabbits injected with soluble BSA. A soluble carbodiimide was used to couple BSA to either heterologous (goat) or homologous erythrocytes. In the latter case, background activity was completely eliminated. Optimal concentrations of BSA and coupling agent are described. The effects on plaque production of incubation time and erythrocyte concentration are discussed. Ammonium chloride was used to remove rabbit erythrocytes, which obscured plaque formation, from spleen suspensions. While incubation with complement (direct assay) detected hemolytically efficient antibody, addition of anti-RγG to the complement (indirect assay) was required to develop plaques due to hemolytically inefficient antibody. Antiglobulin produced in some species inhibited rather than enhanced plaque production, and electrophoretic fractionation of these antisera yielded a predominant blocking fraction and an amplifying fraction. Antisera produced in squirrel monkeys provided consistent amplification without fractionation.
Heterogeneity of Rabbit Anti-Ovalbumin Antibodies Sensitizing Human, Guinea Pig and Rabbit Skin
Authors: Richerson Hal B. , Cheng H. F. , Seebohm Paul M.
Published: December 2022

Three groups of rabbits were injected with five-times recrystallized ovalbumin. One group received soluble ovalbumin, the second alumprecipitated ovalbumin, and the third ovalbumin in complete Freund's adjuvant. The developmental courses of human, guinea pig and rabbit skin-sensitizing antibodies were followed, and representative sera were fractionated by electrophoresis and by Sephadex G 200. The effect of heating at 56°C for 4 hr was also studied.

Rabbit anti-ovalbumin antisera are capable of sensitizing human, guinea pig and rabbit skin. A different antibody sensitizes the skin of each of the three species.

Human SSA was best stimulated by antigen in Freund's adjuvant, was first detectable at 3 to 4 weeks and peaked at 10 to 12 weeks. It is only alightly affected by heat, is fast-moving electrophoretically and elutes with the ascending 7 S peak from Sephadex G 200.

Guinea pig PCA antibody was produced effectively by each immunization technique used, appeared early and continued at high titer for several months. It is heat stable, is slow-moving electrophoretically and coincides with the 7 S peak from Sephadex G 200.

Rabbit homologous PCA antibody was best stimulated by alum-precipitated ovalbumin, was demonstrable at 2 weeks and was usually sustained for several months. This antibody is affected but not destroyed by heat, is fast-moving electrophoretically, and elutes with the ascending 7 S peak from Sephadex G 200.

The GPPCA appears to coincide with the IgG immunoglobulin class, confirming the results of others (12, 13). The immunoglobulin class or classes of HSSA and RaPCA antibodies have not been identified in these studies, but are fast-moving electrophoretically and are perhaps slightly larger in size than IgG immunoglobulins.

In this system, the P-K reaction is not comparable to PCA in the guinea pig.
Antigenic Determinants of Human IgM Requiring Interaction of µ- and κ-Chains
Authors: Miller Edward J. , Terry William D.
Published: December 2022

An antigenic determinant of human κ type IgM proteins which can be detected only when µ and κ polypeptide chains are combined has been investigated. Precipitin in gel and hemagglutination techniques demonstrated that two antisera, which were specific for IgM, detected antigenic determinants present only on IgM preoteins of κ type

Studies were performed to determine whether the antigenic specificity was a characteristic of κ chains or µ chains, or whether both were required. The IgM κ type specificity was retained by the γm8 subunit as well as by enzymatically produced Fab fragments. Antigenic reactivity was not detected in either the heavy or light chains isolated from κ and λ type IgM. The antigenic specificity was demonstrable, however, in reduced, dissociated and recombined molecules consisting of µ chains and κ chains. Expression of this configurational antigenic determinant of κ type IgM proteins, therefore, requires non-covalent interactions between portions of µ chains and κ chains.
Interactions of Sensitized Lymphoid Cells and Homologous Target Cells in Tissue Culture and in Grafts: an Electron Microscopic and Immunofluorescence Study
Authors: Weiss Leon
Published: December 2022

The interactions in tissue culture and in graft of sensitized BALB/c spleen cells and homologous L929 fibroblasts have been studied by light microscopy, electron microscopy and immunofluorescence. The system is slightly modified from that introduced by Rosenau and Moon (2–3).

Spleen cells become adherent to L cells. While erythrocytes, granulocytes, lymphocytes and monocytes and other cell types in the spleen cell suspension adhere to the L cells, the population of initially adherent cells is effectively restricted to plasma cells and lymphocytes (11). Lymphocytes are predominantly the adherent cells at the outset, but plasma cells increase to about 50% of adherent cells in 24 to 48 hr. The peak in plasma cell number coincides with the period of rapid lysis of L cells. Lymphocytes containing polyribosomes, numerous mitochondra, and segments of granular endoplasmic reticulum are frequently observed. Cells containing globulin are revealed by immunofluorescence. Such cells increase in a manner parallel to that of plasma cells. Most immunofluorescent cells, as viewed under phase contrast, moreover, appear to be plasma cells.

I advance the hypothesis that in this system, which is representative of the homograft reaction and perhaps of other instances attributed to delayed hypersensitivity, a class of lymphocytes becomes plasma cells. The plasma cells are secretory cells. They secrete a type or types of antibody or toxin that may itself damage the L cells. But complement may also be produced by adherent plasma cells and, in conjunction with the antibody or toxin, may cause lysis of the homologous cells.
Hereditary Deficiency of the Second Component of Complement in Man: An Immunochemical Study
Authors: Klemperer Martin R.
Published: December 2022

The sera of four homozygous C′2 deficient individuals and 15 heterozygous C′2 deficient individuals in two kindreds were investigated by immunochemical techniques. The decrease in functional C′2 activity in these sera was found to correlate with a decrease in the amount of a serum β globulin which is presumably C′2. Individuals homozygous for the C′2 deficiency lack immunochemically measurable serum C′2. Heterozygous C′2 deficient individuals have 30% to 60% of normal C′2.
Two Antigenic Subtypes of Human Lambda Immunoglobulin Chains
Authors: Tischendorf Frank W. , Osserman Elliott F.
Published: December 2022

Two subtypes St(+) and St(-) of human lambda polypeptide chains were distinguished by immunochemical tests. Five of 27 Bence Jones proteins contained antigenic determinants that were not detected in the other 22 proteins. The distinctive antigenic sites were demonstrated with a heterologous rabbit antiserum produced to a lambda Bence Jones protein. The St(+) lambda chain subtype is present in light chains from pooled normal human immunoglobulin G.

The results of comparative tryptic peptide mapping suggest that the specific St(+) determinants are localized on the variable (N-terminal) half of the lambda chains. St(+) lambda chains have been found to be Oz(-) (arginine residue at position 190 of the C-terminal half of the chain). Twenty-three per cent of the St(-) proteins were Oz(+) (lysine residue at position 190) and 77% of the St(-) proteins were Oz(-).
Effect of Antilymphocytic Serum on Rat Lymphocytes
Authors: Tyler Ruth W. , Everett N. B. , Schwarz M. Roy
Published: December 2022

The results of this radioautographic study demonstrated that ALS destroys large numbers of the long-lived small lymphocytes. Morphologic evidence of destruction was apparent in measenteric lymph nodes and was reflected by reduced numbers of long-lived cells in the thoracic duct lymph. In addition, this agent stimulated some long-lived small lymphocytes to transform into large blast cells. Little, if any, change was noted in the cell content of the thymus and bone marrow, tissues known to contain large numbers of short-lived small lymphocytes. A maximum reduction in the number of long-lived cells was seen by 24 hr after the first injection of ALS and subsequent daily injections for 7 days did not appear to enhance the effect. Although blood lymphocyte levels returned to normal within 6 weeks after cessation of ALS treatment, the number of small lymphocytes in TDL reached only 50% of the control levels. Throughout the experiments normal numbers of short-lived small lymphocytes continued to appear in the TDL of ALS-treated rats. The results are discussed with respect to the immunosuppressive action of ALS.
Autologous Immune Complex Glomerulonephritis
Authors: Glassock Richard J. , Watson J. Ian , Edgington T. S. , Dixon Frank J.
Published: December 2022

Transfer of lymphoid cells from rats with autologous immune complex nephritis induced by intraperitoneal immunization with renal tubular antigen consistently failed to produce disease in normal rats of the same inbred strain. On the contrary, transfer of isologous lymphoid cells from rats with disease induced by intracutaneous and footpad immunization produced a similar, although clinically mild disease, which appeared only after a latent period equal to that needed for the active induction of disease. These observations and those concerning the nature of parabiotic transfer of this disease are consistent with but do not conclusively prove the concept that active immunization of the cell recipient by transferred antigen may play a role in the transfer phenomenon along with any antibody which may be formed by the transferred cells.
Facilitatory and Inhibitory Effects of Phytohemagglutinin on Experimental Allergic Encephalomyelitis in Guinea Pigs
Authors: Shaw Cheng-Mei
Published: December 2022

The in vivo effects of phytohemagglutinin (PHA) on experimental allergic encephalomyelitis (EAE) were studied in guinea pigs: EAE was facilitated when PHA was given 1 week preceding the encephalitogenic sensitization.EAE was suppressed when PHA was given following the encephalitogenic sensitization.The inhibitory effect of multiple post-treatments with PHA was greater than those of a single injection, but multiple pre-treatments did not enhance the facilitatory effect obtained by a single pre-treatment.Subcutaneous injection of PHA stimulated the lymph nodes in vivo, increasing their size and weight and developing marked cytoplasmic pyroninophilia in the cells, first those in the medulla and later those extending into the cortex.

The quite variable effects of PHA on other immune responses in vivo reported by other investigators may be explained by the histologic evidence that PHA affects lymphocytes predominantly. Thus, PHA may not exert so much influence on humoral antibody production as on delayed-type hypersensitivity, such as is responsible for EAE.
A Neutrophil Chemotactic Factor from Human C′5
Authors: Ward Peter A. , Newman Larry J.
Published: December 2022

The action of trypsin on the fifth component of human complement (C′5) results in the appearance of a chemotactic factor for rabbit neutrophils. The generation of this activity is dependent upon the amount of C′5, the duration of incubation and to a lesser degree, the amount of trypsin. On the basis of analysis in density gradient ultracentrifugation and by gel filtration, the chemotactic factor appears to be a cleavage product of C′5 with an estimated molecular weight of 8500. Trypsin appears to be less effective in generating a chemotactic factor from the third component of human complement (C′3).

Chemotactic activity can be also demonstrated in the supernatant from sensitized sheep erythrocytes containing the first four reacting complement components and incubated with sufficient amounts of C′5. In this case, the chemotactic factor sediments very slowly (like the marker cytochrome c) in a sucrose density gradient.

The relationship between the chemotactically active C′5 fragment and F(a) C′5 which has anaphylatoxin activity is not presently known.

These studies document the second fragmentation product of low molecular weight from the complement system possessing chemotactic activity.
Studies on Human Antibodies
Authors: Dorner Marianne M. , Yount William J. , Kabat Elvin A.
Published: December 2022

Six antibodies produced in one individual and anti-dextran formed in different individuals were compared with myeloma globulins in acrylamide gel electrophoresis. They were examined in their intact form as well as after reduction and alkylation. The findings indicate that antibodies and myeloma globulins represent fairly highly selected populations relative to normal γG immunoglobulin, with respect to their banding in acrylamide gel.
Isoantigens Available to Circulating Antibodies
Authors: Witz Isaac , Kaliss Nathan , Pressman David
Published: December 2022

Further studies on the availability of mouse DBA/2 isoantigens to circulating antibodies were made. Isoantibodies isolated from multiparous C57BL females (by DBA/2 males) exhibited in vivo localization in tissues of DBA/2 mice. A drastic reduction in the size of spleen by irradiation did not decrease the fraction of injected isoantibody that was fixed to this organ. Furthermore, the spleens of young mice, although rather small in size, fixed a maximal fraction of the injected isoantibody. Specific fixation of isoantibody by the livers of mice pretreated with immunosuppressive antisera was lower than fixation by the livers of mice treated with normal serum. This may be due either to competition for antigenic sites or to a specific action of the immunosuppressive antibody.
Stimulation of Immunoglobulin and Anamnestic Antibody in Vitro
Authors: Yashphe Diane J. , Coons Albert H.
Published: December 2022

Fragments of lymph node from previously immunized rabbits were secondarily stimulated in vitro and placed under tissue culture conditions with the addition of 14C-leucine to the culture media. Antibody synthesis as determined by incorporation of leucine began earlier than that detectable by passive hemagglutination. It was found that in such cultures the secondary antibody response was superimposed on a steady synthesis of immunoglobulin, possibly antibody directed against other antigens.
A Refractory Period in Adoptive Immunization Following Secondary Stimulation of Mice
Authors: Twarog Frank J. , Rose Noel R.
Published: December 2022

A refractory period of antibody transfer between 8 and 20 days following a secondary stimulation of mice with BSA has been observed. During this period, although donor animals have very high titers of BSA antibody, spleen and lymph node tissue was incapable of producing antibody when transferred to syngeneic recipients. Cells with immunologic memory, however, were present in the transferred tissue as attested by anamnestic responses elicited in the recipient animals with sub-immunogenic doses of BSA. This refractory period was related to the dose of antigen used in the booster injection.
Immunologic Response to Altered Encephalitogenic Protein in Guinea Pigs
Authors: Swanborg Robert H.
Published: December 2022

The immunologic and pathogenetic properties of diazonium derivatives of a purified highly encephalitogenic basic protein (BP) of rabbit origin were investigated. Ars-Sulf-BP, containing three to four azo groups per molecule, was no longer basic by electrophoretic analysis, although the ultracentrifugal characteristics of the original BP were retained.

Guinea pigs immunized with Ars-Sulf-BP did not develop EAE, although delayed hypersensitivitity to BP could be demonstrated. Similar results were obtained when BP was conjugated with the cationic R4N group.

Guinea pigs immunized with Ars-Sulf-BP produced antibodies which appeared to be specific for a determinant larger in size than the haptenic group.
Anti-Dengue Immunoglobulins and Serum β1 c/a Globulin Levels in Dengue Shock Syndrome
Authors: Russell Philip K. , Intavivat Anong , Kanchanapilant Sopana
Published: December 2022

The anti-dengue immunoglobulins present in the sera of 12 children with dengue shock syndrome were studied by immunochemical and serologic methods. IgG antibody was shown to have HI, CF and N activity against dengue viruses. No anti-dengue antibody activity was attributable to IgA or IgM in these patients, all of whom had secondary dengue infections. A depression in serum β1 c/a globulin levels was observed in 23 dengue shock syndrome patients during the shock phase of the illness. Normal levels were found following convalescence. The significance of these observations with respect to possible immunologic mechanisms involved in the pathogenesis of the dengue shock syndrome is discussed.
Immunofluorescence Assay for Antigen and Antibody in Lactic Dehydrogenase Virus Infection of Mice
Authors: Porter David D. , Porter Helen G. , Deerhake Beverly B.
Published: December 2022

An immunofluorescence method to demonstrate infected cells and antibody was developed for lactic dehydrogenase virus infection of mice. Macrophages in the spleen and Kupffer cells in the liver contained viral antigen between 18 and 36 hr post-infection. Mice developed an antibody response to viral antigen by 6 days post-infection.
The in vitro Release of Leukocyte Histamine by Staphylococcal Antigens
Authors: Martin R. Russell , White Arthur
Published: December 2022

Leukocyte histamine was released in vitro by crude staphylococcal antigen supernatants containing protein antigen A or by electrophoretically separated protein antigen A. Leukocyte histamine was not released by similar antigen supernatants from a strain which did not contain protein antigen A, nor by teichoic acids or teichoic acid-mucopeptide complexes. Pre-incubation of antigens with sera containing antigen A precipitins consistently blocked leukocyte histamine release.
Studies on Cryoprecipitation
Authors: Saha Anil , Sambury Sheldon , Smart Kenwyn , Heiner Douglas C. , Sargent Austin U. , Rose Bram
Published: December 2022

Cryoglobulin obtained from a patient suffering from idiopathic cryoglobulinemia was characterized immunochemically. This cryoglobulin (2.7 g/100 ml plasma) was produced by the patient in addition to the normal level of major serum proteins and immunoglobulins (IgG, IgA, IgM and IgD). It possesses a λ-type light polypeptide chain and a γ1-type heavy chain with Gm 4 characteristics. Treatment of the cryoglobulin with papain produced Fab and Fc fragments which individually or collectively showed no cryoprecipitation at 3°C. In contrast, the native cryoglobulin molecule formed a gel matrix when the temperature was reduced below 37°C. Rabbit antisera to this cryoglobulin possessed cryoglobulin-specific antibody determinants which were directed toward the Fd pieces of the Fab fragment. The Fab fragments displayed a dimermonomer equilibrium depending on the hydrogen ion concentration of the media.
Chromatographic Studies on Spleen and Lymph Node Extracts with Antibody Activity
Authors: Tsuji Frederick I. , Davis David L. , Donald David H.
Published: December 2022

Rabbits were immunized with the enzyme Cypridina luciferase and the antiluciferase activity in spleen and lymph node extracts was studied chromatographically. The extracts were separated into major fractions by differential centrifugation and into subfractions by gel exclusion chromatography. Only two types of soluble antiluciferase activity were found. Based on elution from Sephadex G 200 columns, the first type of activity appeared to be due to immunoglobulin-G (IgG). The second type possessed a molecular weight between 50,000 and 70,000 and resembled Porter's fragments I and II. Significant amounts of luciferase were associated with the high molecular weight fraction of lymph nodes.
Hapten-Specific Insoluble Immunoabsorbents Prepared from Hide Powder (Collagen)
Authors: Levine Bernard B. , Levytska Vera
Published: December 2022

Hapten-specific immunoabsorbents using bovine hide powder (an inexpensive material consisting largely of collagen) as the matrix were prepared by coupling it with hapten, and were used to isolate antibodies from rabbit anti-hapten sera. Both benzylpenicilloyl (BPO) and dinitrophenyl (DNP) hide powder had high absorptive capacities for antibody. Absorption and elution were specific. Antibody recoveries ranged from 75% to 100% when eluted with hapten. Antibody purity was high; specific precipitability was 95% to 100% for BPO and 45% to 86% for DNP. DNP antibody preparations contained hapten as an impurity, which appears to account for the low precipitability. BPO antibody preparations were virtually free of hapten. The immunoabsorbents are conveniently prepared, economical and stable. Hide powder may have wide applicability as an insoluble matrix for a variety of biologically important small and large molecules.
Relationship Between Viremia and Interferon Production of Japanese Encephalitis Virus
Authors: Rokutanda Hinae K.
Published: December 2022

Viremia with JE virus and its relationship to pathogenicity were studied in mice. The degree of viremia and susceptibility to peripheral infection decreased with age. Mice became markedly resistant to JE virus infection between 1 and 2 weeks of age. A viremia-positive (V+) variant was isolated by rapid blood-to-blood passages of the Mukai strain in 1-week-old mice. This strain causes the highest viremia with JE virus ever reported in 3-week-old mice. The maximal titer was about 103 PFU/ml of blood when 30 PFU were inoculated subcutaneously. In general, viremia with JE virus was not detected so easily and its titer was not so high when the virus was inoculated subcutaneously in 3-week-old mice. There was a proportional relationship between the occurrence of viremia and pathogenicity. The V+ variant showed 100-fold higher pathogenicity in peripheral infections and 10-fold higher in intracerebral infection than the wild type (V-) virus in 3-week-old mice.

The causal relationship between the occurrence of viremia and the production of circulating interferon was studied. V- virus produced more interferon in the blood of mice than the V+ virus, whereas the titer of viremia was higher with the V+ variant than with V- virus. So the occurrence of viremia was regulated by the ability of the virus to produce circulating interferon. No difference was found in sensitivity to interferon between V+ and V- virus.
Mycoplasmacidal Effect of Polymorphonuclear Leukocyte Extract
Authors: Dajani Adnan S. , Ayoub Elia M.
Published: December 2022

The effect of leukocyte extract on the viability of certain human Mycoplasma was investigated. Leukocytes were obtained from venous blood of human donors or from glycogen-induced peritoneal exudate of rabbits. The cells were suspended in citrate buffer at various pH values and the extract was prepared by repeated quick-freezing and thawing. The supernatant of these preparations showed a marked mycoplasmacidal effect on M. pneumoniae, M. salivarium and M. fermentans. Fractionation with ammonium sulfate revealed that the activity of this extract resides in the fraction obtained at 40% to 60% saturation of ammonium sulfate. This mycoplasmacidal factor exhibits optimal activity at about pH 4.7, is non-dialyzable and is relatively heat resistant.
DNA-Binding Assay for Detection of Anti-DNA Antibodies in NZB/NZW F1 Mice
Authors: Steinberg Alfred D. , Pincus Theodore , Talal Norman
Published: December 2022

NZB/NZW F1 mice (B/W) spontaneously develop an autoimmune disease resembling human systemic lupus erythematosus (SLE) (1, 2). This includes LE cell and anti-nuclear factor formation, severe glomerulonephritis, and uremia. The serologic and histologic abnormalities appear earlier in the female B/W mice who generally die of nephritis by 10 months of age. The renal disease is considered a soluble immune complex type of nephritis (3). DNA, anti-DNA antibodies and complement have been demonstrated in the glomeruli of these mice (3) as well as in patients with SLE (4).

Previous studies demonstrating serum anti-DNA antibodies in B/W mice have employed the techniques of immunofluorescence (3, 5), immuno-diffusion and complement fixation (3). These indirect methods are limited by variable patterns of fluorescent staining, non-precipitating antibody and anti-complementary activity. Recently a sensitive assay measuring the direct binding of radioactive DNA by immunoglobulin has been described for humans with SLE (6, 7).
Delayed Hypersensitivity
Authors: Lipsmeyer Eleanor A. , Kantor Fred S.
Published: December 2022

Pre-exposure of sensitized guinea pig peritoneal exudate cells to antigen renders them incapable of migration inhibition in capillary tubes. However, pre-exposure to the same antigen did not affect migration inhibition of heterologously sensitized cells. After a population of cells sensitized to multiple antigens was pre-exposed to one of them, none of the antigens was capable of inhibiting migration in the capillary tube system.
Antigen-Induced Cell Proliferation in Cultures from Antibody-Deficient Chickens
Authors: Meuwissen H. J. , Van Alten P. J. , Good R. A.
Published: December 2022

Bursectomized and irradiated, hypogammaglobulinemic chickens and irradiated controls were immunized with killed Brucella abortus. Cultures of splenic cells from these animals were subsequently stimulated with the same antigen. Hypogammaglobulinemic chickens did not produce antibody in vivo, but the uptake of tritiated thymidine after antigenic stimulation in vitro was unimpaired.

The results indicate that lymphoid cell lines dependent on the bursa of Fabricius for development to immunoglobulin-producing cells are not necessary for a vigorous response to Brucella antigens in vitro.
An Isotopic Antiglobulin Technique for Measuring Antibodies to Cell-Surface Antigens
Authors: Sparks F. C. , Ting C. C. , Hammond W. G. , Herberman R. B.
Published: December 2022

The isotopic antiglobulin technique of Harder and McKhann has been modified and the sensitivity increased. The method was shown to be immunologically specific and able to detect isoantibodies after first-set skin grafts which were not detectable by the cytotoxicity technique. Antibodies to different H-2 antigens were detected in the sera of several inbred mouse strains following immunization with spleen cells or skin grafts. In several of these antisera, antibodies were detected at serum dilutions which gave negative results by the cytotoxicity technique. Several variables of the technique were investigated and the number of washing procedures after each incubation was found to have the greatest effect on increasing the sensitivity of the test.