bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of Thymectomized, Irradiated Rats to Study Immunogenicity of Bovine λ Globulin
Authors: Williams R. Michael , Waksman Byron H.
Published: December 2022

Adult Lewis rats, thymectomized, x-irradiated, and restored with syngeneic bone marrow, developed immunologic memory when 4 to 6 × 108 normal Lewis lymph node cells were injected before or up to 5 days after footpad injection of 1.0 mg of “crude” BGG in saline. Increasing the time interval from 10 to 30 days between x-irradiation plus bone marrow repopulation and the injection of lymph node cells plus primary antigen resulted in increased priming. Injection of lymph node cells up to 40 days before primary immunization had little effect on memory production. No priming could be demonstrated when lymph node cells were given 10 or more days after primary antigen.
The Effects of Ionic Strength, Temperature, and Antibody Class and Avidity on Fixation and Transfer of the First Component of Complement
Authors: Linscott William D.
Published: December 2022

Over a wide range of ionic strengths the first component of complement (C′1a) was found to be readily eluted from IgG-sensitized cells at temperatures above 0°C. Even at 0°, loss of C′1a was high at ionic strengths appreciably above 0.037. With IgM Ab the effects of ionic strength were similar, but those of temperature were reversed: C′1a was increasingly better retained on cells as the temperature increased from 0° to 37°C. Conditions for reaching optimal C′1a activity with both classes of antibody (Ab) were determined.

The optimal ionic strength for transfer of C′1a from EAC′1a to EAC′4 (red cells sensitized by Ab and the first and fourth components of complement, respectively) was found to be approximately 0.115 for both IgG and IgM Ab. When all conditions of C′1a fixation, washing and transfer were optimal, essentially 100% of the C′1a originally bound to sensitized cells could be accounted for by transfer to EAC′4, regardless of the class of Ab used. Transfer of C1a from IgG-sensitized cells was appreciably better at 30°C than at 0°, but with IgM Ab the converse was true. Kinetic studies showed that transfer was essentially complete within 5 to 15 min.

C′1a dissociated less readily from EAC′1a, 4 than from EAC′1a, indicating that cell-bound C′4 contributes to the bond between C′1a and its site of attachment.

In contrast to the situation with IgM Abs, with IgG it was not possible to reach a saturation level of C′1a; with each increase in amount of C′1a added, there was an increase in the number of SAC′1a generated, even when the ratio of C′1a:SAC′1a exceeded 100:1.
Immunochemical Relationships of Certain Streptococcal Group and Type Polysaccharides to Pneumococcal Capsular Antigens
Authors: Heidelberger Michael , Willers Jan M. N. , Michel Marc F.
Published: December 2022

The quantitative data on cross-reactions of streptococcal group-like and group F polysaccharides permit these conclusions:

z1 apparently contains non-reducing end-groups of L-rhamnose; z2 precipitates antipneumococcal type VII serum relatively independently of the type II substance occurring in the same cell but has groupings in common with TIV; z3 appears to contain 1,3-linked L-rhamnose; z4 overlaps markedly with TIV and z6 in anti-PnVII and reacts as if it contained end-groups of D-galactose.

Type I polysaccharide overlaps somewhat with z1 in anti-PnVII and with TIII in anti-PnX; galactose is probably involved. Precipitation of anti-PnVII with TII removes some of the antibody reactive with z2; after both are removed there is no further precipitation with z1. Possibly PnSVII and z1, z2 and TII all contain 1,2-linked L-rhamnose or a few non-reducing end-groups of this sugar. The reactions of TII in anti-PnVIII and -XIX show for the first time that the long-known cross-reaction between PnVIII and XIX is due to multiple residues of 1,4-linked D-glucose.

Like TI, TIII cross-reacts in anti-PnX and also precipitates anti-PnXIII, XIV and XX, the last not being due to glycogen-like linkages of D-glucose. The strong reactions of TIV with anti-PnVII and XVI are accounted for by multiples of non-reducing end-groups of β-D-galactose, as is perhaps also the heavy precipitation in anti-PnXVII. TIV also precipitates anti-PnVIII and -XIX, at least partly for the same reason as TII. z6 shows almost complete overlapping with z1 in anti-PnVII and -XXIII.
A New Allotypic Antigen of Rabbit Colostral γA Immunoglobulin
Authors: Masuda Tohru , Kuribayashi Kagemasa , Hanaoka Masao
Published: December 2022

Rabbit isoantiserum to colostral γA of a rabbit with genotype a2a2-b4b4 reacted with some normal rabbit sera and colostrums. The reaction was due to antigenic specificity present only on α chain, but not on the Fab part of γA and γ chain. This determinant was identified as a new rabbit γA allotype and was localized on the Fc part. This new allotype was named c1, and its hypothetical controlling gene locus, “c.”

Among 70 rabbits tested, the sera and colostrums of 11 showed c1 specificity. The c1 marker was expressed not only in rabbits of a genotype a2a2-b4b4, but also in rabbits lacking in a2 or b4 specificity (for example, a1a3-b4b5).

Allotypes a1 and a3, in addition to b4 and b5, were found in colostral γA molecules and their subfragments, Fab.
The Response of Long- and Short-Lived Small Lymphocytes of the Rat to Pokeweed Mitogen
Authors: Stayner Larry , Schwarz M. Roy
Published: December 2022

Small lymphocytes of the thoracic duct lymph, mesenteric lymph node, spleen and thymus from Lewis and Wistar rats were exposed to PWM in vitro. The results of radioautographic studies reveal that both the long- and short-lived small lymphocytes transformed in the presence of PWM, but the short-lived populations of the thoracic duct lymph, mesenteric lymph node and spleen were more responsive. In contrast, the short-lived cells of the thymus transformed less than the long-lived small lymphocytes of the same organ. The responsiveness of short-lived small lymphocytes from a given tissue reflected the number of cells from this tissue required to initiate graft-versus-host reactions.
Preparation of Fluorescent Antibody Reagents Monospecific to Light Chains of Human Immunoglobulin
Authors: Takahashi Morinobu , Yagi Yasuo , Pressman David
Published: December 2022

Anti-light chain antibodies that react well with separated light chains and intact immunoglobulins of human beings could be readily produced in rabbits and goats by immunization with the Fab fragment of normal human IgG. Antibodies against light chains of κ or λ type were specifically purified from rabbit anti-normal Fab sera and rendered monospecific by the exclusive use of solid immunoadsorbents coupled with appropriate antigens. The anti-κ or anti-λ antibodies thus prepared were conjugated with fluorescein and tetramethylrhodamine and used for the study of immunoglobulin production in individual cells of hematopoietic origin in established culture.
Communications
Authors: Linscott William D.
Published: December 2022

When EAC′1 (erythrocytes sensitized by antibody and the first component of complement) are prepared with an amount of IgG antibody (Ab) just sufficient to produce about 100 C′1-fixing sites/cell, the C′1 can be quantitatively transferred from these cells in a short time at 30°C at ionic strength (µ) = 0.063. If 50 times as much IgG Ab is used, but C′1 is limited to produce about 100 or fewer SAC′1/cell, then almost no C′1 transfer occurs at 30°C, µ = 0.063, but much of the C′1 will transfer at µ = 0.14. It is suggested that C′1 bound to IgG dimers transfers readily at µ = 0.063, while C′1 bound to IgG trimers transfers readily only at µ = 0.14 or higher, and C′1 bound to IgG tetramers may fail to transfer altogether. With IgM Ab, a slight similar effect was noted, to the extent that for equivalent amounts of C′1 transfer it was necessary to increase the ionic strength as larger amounts of IgM were used to sensitize erythrocytes.
Density of BAI Strain A Avian Leukosis Virus-Antibody Complexes Relative to Antigen-Antibody Ratio
Authors: Fritz Robert B. , Beard J. W.
Published: December 2022

Immune complexes of differing antibody-virus ratios were prepared with rabbit anti-BAI strain A avian leukosis virus and the homologous antigen by means of the quantitative precipitin reaction. These complexes were centrifuged to their equilibrium densities on potassium tartrate gradients and the results were compared with the analogous properties of free virus and normal rabbit γ globulin centrifuged under the same conditions. The equilibrium density of the complexes was greater than that of the free virus and was directly proportional to the antibody-virus ratio. Normal rabbit globulin was not sedimented significantly under the same experimental conditions.
Cytophilic Antibody in Heterologous Antilymphocytic Serum: Its Presence, Absorption and Possible Biologic Significance
Authors: Gill P. G.
Published: December 2022

Recent studies on the mode of action of antilymphocytic serum (ALS) have suggested that its primary site of action is on the thymus-derived small lymphocyte (1, 2). It has also been suggested that these cells are eliminated rather than temporarily inactivated (3) and that an important part of this process is opsonization followed by phagocytosis (2). Greaves et al. (4) recently showed that there was a significant correlation between the immunosuppressive potency of ALS, and its opsonic titer as estimated by a technique involving the adherence of opsonized mouse lymphocytes to mouse peritoneal macrophages in monolayer culture.

The present communication describes macrophage-cytophilic antibody in rabbit anti-mouse-lymphocytic serum (RAMLS). This may well be of significance in relation to the work cited above, as cytophilia has been defined as the property of opsonizing antibody which provides the receptors that permit binding of antigen to the macrophage cell membrane in preparation for phagocytosis (5).
Purification of the First Component (C′1) of Complement by Zonal Ultracentrifugation and of the Second (C′2) by Electrofocusing
Authors: Colten H. R. , Borsos T. , Bond H. E. , Rapp H. J.
Published: December 2022

The zonal centrifuge has been used to purify viruses, subcellular organelles and macromolecules of biologic interest. In this report we describe a preparative zonal ultracentrifugation method for the separation from serum of chemically and functionally highly purified first component (C′1) of guinea pig and human complement. The observation that the sedimentation rate of C′1 varies inversely with the ionic strength of the medium made the development of this purification procedure possible. At an ionic strength of 0.3 or greater the sedimentation constant of C′1 is about 4 and at an ionic strength of 0.15 or less the sedimentation constant of C′1 is about 18. The purification procedure consists of a two-cycle centrifugation: the first at µ = 0.75 separates C′1 from 19 S serum proteins; the cycle at µ = 0.065 separates C′1a, now an 18 S molecule, from the smaller molecules.
Studies on the Seventh Component (C′7) of Complement
Authors: Mayumi M. , Shimada K. , Sekine T.
Published: December 2022

From experiments with sucrose density gradient ultracentrifugation and Sephadex G-200 gel filtration the molecular weights of guinea pig C′7, human C′7 and rabbit C′7 were estimated to be 120,000, 130,000 and 130,000, respectively. In zone electrophoresis on Geon or agarose, C′7 of human, guinea pig and rabbit serum migrated, in this order, in the β globulin region.

The C′7 of the three species reacted well with EAC′1–6 prepared with guinea pig components. From quantitative analysis of C′7 site formation, the dose response curve yielded a straight line when this reaction step was analyzed with the aid of Mayer's one-hit theory. Guinea pig C′7 reacted with EAC′1–6 at an ionic strength of µ = 0.15 as well as at an ionic strength of µ = 0.74. C′7 activity was shown to disappear from the fluid phase after the reaction with EAC′1–6.

Guinea pig antiserum to rabbit C′7 inhibited the hemolytic activity of human and rabbit C′7 but not that of guinea pig C′7.
Immunochemical Studies on Blood Groups
Authors: Moreno Carlos , Kabat Elvin A.
Published: December 2022

Humans immunized with blood group A substances from hog gastric mucosa may produce antibody specific for a terminal non-reducing α-D-GNAc residue. This antibody is unrelated to human anti-A.
The Serum Bactericidal Reaction. III. Antibody and Complement Requirements for Killing a Rough Escherichia Coli
Authors: Goldman John N. , Ruddy Shaun , Austen K. Frank , Feingold David S.
Published: December 2022

The serum bactericidal reaction was studied using the rough E. coli 200P and a system of serum reagents made selectively deficient in various complement components. In replacement experiments functionally pure individual components were added and requirements for C1, C4 and C2, and C9 as well as antibody were shown. Cellular intermediates BAC1 and BAC142 were demonstrated. The results suggest that the entire complement system is required to kill bacteria.

Note Added in Proof. Inoue et al. (Inoue, K., Yonemasu, K., Takamizawa, A. and Amano, T. Biken J., 11: 203, 1968) have recently reported that BAC142 cells, C3, 5, 6, 7, 8, 9 and lysozyme were required for immune bacteriolysis of a rough strain of E. coli. The complement components used were purified from guinea pig serum. Results were based on observations of spheroplast formation: with the complete system about 25% of the rod-shaped bacteria were converted to spheroplasts.
Immunoglobulin Synthesis by Cultured Mouse Myeloma Cells
Authors: Namba Yuziro , Hanaoka Masao
Published: December 2022

Murine myeloma cells (MOPC-31B) were established in long term culture and retained the capacity to produce and secrete IgG myeloma proteins. The mechanism of synthesis of myeloma protein was analyzed at the cellular and subcellular levels, and the following results were obtained. Light and heavy chains are separately synthesized on different polysomes, 200 S and 280 to 300 S with 12 S and 22 S messenger RNA, respectively. Synthesized heavy chains form H-L complexes with free light chains, which are maintained as a large pool in the cytoplasm, then H2-L2 complexes are formed and secreted after the attachment of carbohydrate in considerable amount.
The Influence of Heredity and Environment on Human Immunoglobulin Levels
Authors: Allansmith M. , McClellan B. , Butterworth M.
Published: December 2022

IgG, IgA, IgM, IgD, kappa and lambda levels were determined in 24 sets of fraternal twins, 21 sets of identical twins, and 15 normal subjects bled weekly for 25 weeks. By statistical comparison of differences among these groups, the influences of heredity and environment on immunoglobulin levels were determined. An hereditary influence was detected on the levels of immunoglobulins IgG, IgA and IgD, and an environmental influence was detected on the levels of immunoglobulins IgA and IgM and on light chains kappa and lambda.
Some Antigenic and Physical Properties of Virus-Like Particles in Sera of Hepatitis Patients
Authors: Barker Lewellys F. , Smith Kendall O. , Gehle Warren D. , Shulman N. Raphael
Published: December 2022

The Australia antigen was detected by Blumberg, Alter and Visnich in 1965 in human sera by formation of a precipitin line in Ouchterlony gel diffusion slides (1). Antibodies used to detect the antigen are found in sera from patients who have received multiple transfusions The antigen is rare in healthy Americans, but common in patients with either infectious or serum hepatitis, patients with Down's syndrome who have anicteric hepatitis, patients with leukemia or leprosy, and apparently healthy persons in certain tropical populations (2). Bayer, Blumberg and Werner described 19 to 21 mµ particles in antigen-containing serum fractions from sucrose gradients (3). The particles were aggregated by the same sera that precipitated the antigen. Blumberg and co-workers and Prince have suggested that the antigen is a virus or is intimately associated with a virus that is responsible for hepatitis in man (2–5).
A Graft vs. Host Reaction in a Xenogeneic Combination: The Reaction of Murine Cells in Rats
Authors: Saleh Wasfy , MacLean Lloyd D. , Gordon Julius
Published: December 2022

The injection of viable immunologically competent mouse cells under the kidney capsule of cyclophosphamide-treated rats produces a localized graft vs. host (GVH) reaction, manifested by infiltration and destruction of the kidney cortex. The reaction, shown to be a GVH reaction by cytological analysis, can be inhibited by antisera specific to the cells injected.
A Role of Macrophages in the Stimulation of Immune Responses by Adjuvants
Authors: Unanue Emil R. , Askonas Brigitte A. , Allison Anthony C.
Published: December 2022

Bordetella pertussis vaccine or beryllium sulfate proved to be potent adjuvants in increasing the antibody response to Maia squinado hemocyanin in mice. Antigen-containing macrophages treated with B. pertussis in vitro elicited much higher antibody titers when injected into mice than they did in the absence of the adjuvant. Lymph node cells treated in vitro with adjuvants and injected into irradiated mice gave the same antibody titers as they did in the absence of adjuvants. Hence in the system employed, adjuvants were shown to exert their enhancing effects on antibody-producing cells only after uptake by macrophages. Adjuvant-treated macrophages injected simultaneously with antigen-containing macrophages also led to increases in the immune response; thus adjuvant action did not require the presence of antigen and adjuvant in the same macrophage. The overall catabolism and retention of 131I-labeled hemocyanin in macrophages was not significantly altered in the presence of adjuvants.
Capillary Tube Migration for Detection of Human Delayed Hypersensitivity: Difficulties Encountered with “Buffy Coat” Cells and Tuberculin Antigen
Authors: Kaltreider H. Benfer , Soghor David , Taylor Janie B. , Decker John L.
Published: December 2022

The usefulness of a simple in vitro capillary tube migration system using human buffy coat cells has been investigated in relation to tuberculin skin sensitivity. A lack of reproducibility of the method was demonstrated and the movement of cells out of capillary tubes was found to depend primarily upon gravity. No correlation between cell “migration” and skin sensitivity was found. It is concluded that this convenient technique, in its present form, is not suitable for the detection of delayed hypersensitivity in man.
Assessment of a Bacterial Adherence Colony (BAC) Method for Enumeration of Antibody-Forming Cells to E. Coli Somatic Antigen
Authors: Friedman Herman , Allen Jerry , Landy Maurice
Published: December 2022

The number of individual lymphoid cells producing antibody to E. coli somatic lipopolysaccharides was studied in mice by means of a bacterial adherence colony technique. Cultures of spleen cells with bacteria in nutrient agar plates resulted in formation of adherent micro-colonies which could be differentiated microscopically from the much smaller colonies originating from non-adhering bacteria. The specificity of the antibody-producing cells was attested by their sensitivity to blocking with specific antigen and with anti-globulin serum.

The adherence technique, performed in parallel with the direct complement-dependent bacteriolytic procedure, using a “lawn” of viable E. coli, detected more “background” antibody-forming cells in spleens of non-immunized mice. This difference was of the order of 10-fold.

In immunized mice bacterial colony-forming cells were first discernible about 24 hr after immunization and reached a peak of 100,000 or more in the spleen by the 5th day after antigen atimulation. Maximal responses were elicited by 10 to 50 µg of antigen. The bacteriolytic plaque assay was less sensitive throughout.
Studies on the Radiosensitive Phase of the Antibody Response in Rabbits
Authors: Pribnow James F. , Silverman Myron S.
Published: December 2022

Macrophages from irradiated donors were shown to recovery the ability to induce antibody formation in normal recipients. Recovery took place within approximately 27 days, regardless of whether cells were transferred to recipients 5 days or 20 days after irradiation of donors.

Irradiated recipients of sensitized macrophages formed no antibody during the first 21 days after cell transfer. These rabbits responded to challenge with BGG with what appeared to be a typical secondary response. Recipients of irradiated sensitized cells (750 r) responded in a similar manner. When recipients of sensitized macrophages had received 750 r rather than 450 r, there was neither a primary nor a secondary response.
Effect of Antimacrophage Serum on Antibody Production and Phagocytosis in Mice
Authors: Argyris Bertie F. , Plotkin Diane H.
Published: December 2022

During the last year four reports have appeared in the literature on the effect of antimacrophage serum (AMS) on phagocytosis and antibody production (1–4). There is general agreement that AMS is cytotoxic to macrophages (1–4) and can decrease the phagocytic activity of peritoneal macrophages (1–4), but some investigators find no immunosuppressive effects after AMS treatment (1, 3), and others (2) report immunosuppression only under certain conditions of antigen administration.

In this report we present evidence that AMS, which decreases the phagocytic activity of mouse peritoneal macrophages, also inhibits primary antibody synthesis. The immunosuppressive effect, however, can only be detected after pretreatment with AMS and low doses of antigen.

Rabbit anti-mouse macrophage serum (AMS) was prepared by injecting New Zealand rabbits three times at bi-weekly intervals, i.v., with 108 or 109 peritoneal cells from C3H/HeJ donor mice. Peritoneal cell donors were injected 7 days previously with 3 ml of thioglycollate medium (5).
The Effect of Carbamylation, Amidination and Benzylation on the Ability of Rabbit γG Antibody to Produce Passive Cutaneous Anaphlaxis in Guinea Pig and Rabbit Skin
Authors: Cohen Stanley , Feinman Lawrence , Becker Elmer L.
Published: December 2022

The effect of chemical modification of amino groups of rabbit γ G antibody on its PCA activity in guinea pig and rabbit skin was studied, with particular attention to high degrees of carbamylation which have profound effects on the antibody's ability to fix complement and to aggregate with antigen. It was concluded that there is no evidence at present for the selective inactivation of specific PCA sites on the Fc portion of the antibody molecule by these reagents. Rather, the observed loss of PCA reactivity in rabbit skin could be attributed to the impaired ability of the antibody to fix complement. Loss of PCA activity in guinea pig skin required greater than 90% carbamylation of the antibody molecule and could be explained in terms of the nonspecific effects on the molecule produced by such treatment. It was further observed that the loss in PCA activity under these conditions could be potentiated in a system in which cross-reacting antigen was used.
Effects of Radiation on the Initial and Anamnestic IgM Hemolysin Responses in Rabbits; Antigen Injection After X-Rays
Authors: Taliaferro William H. , Taliaferro Lucy Graves
Published: December 2022

The effects of a wide range of x-rays from 25 to 700 r on the initial and anamnestic IgM hemolysin responses were ascertained in groups of rabbits by giving an intravenous initial injection or reinjection of 108.3 fresh sheep red blood cells (SRBC)/kg rabbit from 4 hr to 2 months (and in some cases 6 months) after irradiation. Sera from the rabbits were titrated by the delicate 50% hemolysis endpoint and graphs of the responses were analyzed with respect to five parameters: length of the latent period, peak titer, length and rate of rise to peak titer and decline from peak titer. The following statistically significant modifications were revealed, as compared to corresponding initial and anamnestic responses in unirradiated control groups. 1) On the one hand, in the initial response, small doses of 25 and 100 r stimulated a) the prolonged production of hemolysin when SRBC were injected from 4 hr to a week thereafter and b) a transient high peak titer when SRBC were injected at 1 month after irradiation. On the other hand, 25 and 100 r did not stimulate the anamnestic response at all, but depressed peak titers when SRBC were given from 7 to 30 days thereafter. 2) 25 and 100 r did not modify the length of the latent period in either response. 3) Large doses of 500 and 700 r equally and intensively depressed both responses when SRBC were given from 4 hr to 2 days thereafter. Such responses were characterized by prolonged latent periods and prolonged hemolysin rises at low rates of rise to depressed peak titers. 4) Recovery from the depressing effects of 500 and 700 r began when SRBC were given 7 days after irradiation, but was more delayed in the anamnestic response than in the initial response.

The foregoing differences between the initial and anamnestic hemolysin responses in rabbits, stimulated with relatively small amounts of Forssman antigen after irradiation, suggest that the immunologically competent initial cells, as compared to the activated memory cells, are more easily stimulated and less sensitive to injury, and that both types of cells are modified to a greater extent during induction than during later stages of proliferation and synthesis.
Immunoenzymology of Amp-Deaminase
Authors: Selig Donald , Chilson Oscar P.
Published: December 2022

The immunologic specificities of crude and partially purified rabbit antisera directed against homogeneous AMP-deaminases from chicken breast muscle and rat skeletal muscle have been investigated. In contrast to a previous report, the results of a comparative study of avian and mammalian enzymes show that the skeletal muscle adenylate deaminases from these organisms are antigenically related. A variety of techniques, including double diffusion in two dimensions, immunoelectrophoresis, and enzyme inhibition, have been used to demonstrate varying degrees of cross-reaction between antibodies and enzymes from different animals. The observation that cross-reactive autoantibodies are produced in response to immunization of rabbits with adenylate deaminases from heterologous (chicken and rat) sources is of particular interest. While the underlying mechanisms are not known, it is clear that these results are not due to the effect of adjuvants or the participation of isoantigens.
Passive Cutaneous Anaphylactic and Schultz-Dale Reactions Elicited in Guinea Pigs by Chemically Univalent Dinitrophenyl-Derivatives of Bacitracin A and Papain
Authors: Mancino Domenico , Tigelaar Robert E. , Ovary Zoltan
Published: December 2022

Passive cutaneous anaphylaxis (PCA) and Schultz-Dale reactions were elicited with mono- and poly-DNP-derivatives of bacitracin A and a mono-DNP-derivative of papain. With high affinity rabbit antibody, the same endpoint titers for PCA could be obtained with tri-DNP-Bac, di-DNP-Bac and DNP-orn-Bac, as well as with bis-DNP-lysine, whereas a significantly lower titer was obtained with DNP-ile-Bac. DNP-his-Bac was ineffective under the conditions used. A significantly higher PCA titer was obtained with highly conjugated DNP-BSA. The relative effectiveness of the DNP-derivatives of bacitracin and of bis-DNP-lysine was correlated with the affinity of the antibody for the appropriate ligands as measured by equilibrium dialysis. With rabbit antibody of lower affinity neither tri-DNP-Bac, DNP-orn-Bac, DNPL-S-pap, nor bis-DNP-lysine were able to elicit PCA reactions. The possibilities that the mono-DNP-derivatives of bacitracin and papain are able to elicit anaphylactic reactions as univalent compounds or rather as functionally multivalent antigens produced either by binding to serum proteins or by molecular aggregation are discussed in the light of the experimental results.
Reaginic-Type of Antibody in Mice Stimulated by Extracts of Bordetella Pertussis
Authors: Clausen C. R. , Munoz J. , Bergman R. K.
Published: December 2022

The Rocky Mountain Laboratory (RML) strain of albino mouse responds to the injection of hens' egg albumin (EA) and soluble extracts of Bordetella pertussis with the production of two homocytotropic antibodies. One of these antibodies (referred to as 2-hr PCA antibody) is a 7S immunoglobulin which does not persist in the skin of mice for a long time and is heat stable and resistant to reduction and alkylation. The other antibody (referred to as 72-hr PCA antibody) is similar to the reaginic antibody of allergic persons, since it is inactivated by heating undiluted sera at 56 C for 30 min and by reduction and alkylation; it remains in the skin for a long time and is not a 7S or 19S immunoglobulin but probably a new type similar to the IgE immunoglobulin of allergic persons. B. pertussis alkaline saline extract (SE) was the best adjuvant for the production of the 72-hr PCA antibody, while Freund's adjuvant was superior in stimulating the 2-hr PCA antibody. Endotoxins from Salmonella minnesota or from B. pertussis were not as effective in stimulating the 72-hr PCA antibody as was the SE.

The production of 72-hr PCA antibody in RML mice persisted for at least 60 days after a single immunization with EA and SE, while the sera of mice immunized with EA in Freund's incomplete adjuvant failed to give 72-hr PCA 40 days after immunization.

The 72-hr PCA antibody was found to be resistant to heating at 56°C for 30 min. if the serum was diluted in saline prior to heating, which indicated that this antibody is inactivated by a mechanism other than heat denaturation.
Isoenzyme Differences between a Human Diploid Cell Line, WI-38, and SV40-Transformed WI-38
Authors: Bartholomew Eleanor M. , Bartholomew William R. , Rose Noel R.
Published: December 2022

Enzymatic differences between diploid and SV40-transformed human fetal lung cell lines were ascertained using enzymo-electrophoresis and enzymo-immunoelectrophoresis in agarose gel. Saline extracts of WI-38 and its transformed counterpart could be differentiated on the basis of lactic dehydrogenase and esterase activities. A cathodal esterase isoenzyme, sensitive to diisopropyl fluorophosphate and resistant to eserine, was found in WI-38, but was depleted at least fivefold in the transformed cell line.
The Binding Reactions of Antibodies Specific for the 2,6-Dintrophenyl Group
Authors: Little J. Russell , Border Wayne , Freidin Ralph
Published: December 2022

Rabbit and guinea pig antibodies specific for the 2,6-DNP group were isolated from sera by specific precipitation and hapten elution. Anti-2,6-DNP-antibodies formed late in the immune response displayed greater cross-reactivity with 2,4-DNP and 2,4,6-TNP ligands than was observed with antibodies obtained at earlier times. Rabbit anti-2,6-DNP molecules were found to have low Qmax values (in the same general range as rabbit anti-2,4,6-TNP molecules). Hypochromic and bathochromic spectral shifts were observed when ε-2,6-DNP-aminocaproate formed complexes with rabbit or guinea pig antibodies or with tryptophan (as the free amino acid) supporting the previous suggestion that one or more tryptophan residues occur in the binding sites of antibody molecules specific for polynitrophenyl determinant groups.
Immunosuppression by Antigen-Associated Human Serum Alphaone Protein
Authors: Whang H. Y. , Chun D. , Neter E.
Published: December 2022

The effects of human serum alpha-one protein on the immune response of the rabbit to two bacterial antigens, one produced by enterobacteriaceae and the other by Gram-positive bacteria, were investigated. It was shown that this protein markedly inhibits the production of circulating antibodies, provided that antigen and alpha-one protein are mixed and incubated prior to injection. Suppression of the immune response does not occur when the two materials are injected separately, albeit simultaneously. Alpha-one protein does not destroy the antigenic determinants of the two antigens. Animals immunized with mixtures of antigen and alpha-one protein are primed. Alpha-one protein is significantly less effective as an antigenassociated immunosuppressant against the common enterobacterial antigen produced by E. coli O14 than it is against the moiety with identical antigenic determinants produced by Salmonella typhimurium.
Differences in Anti-Protein a Activity among IgG Subgroups
Authors: Kronvall Göran , Williams Ralph C.
Published: December 2022

The reaction between staphylococcal protein A and human γG globulin, which is known to be mediated by structures on the Fc part of the heavy chains, was studied qualitatively using 68 isolated γG myeloma globulins. Twenty-one myeloma globulins of subgroups γG-1, γG-2 and γG-4 gave a precipitin reaction with protein A. The 35 remaining meyloma globulins of these subgroups all inhibited this reaction, indicating the presence of protein A-reactive sites. Twelve γG-3 myeloma globulins did not precipitate or inhibit the precipitation; two showed slight inhibition that could be attributed to background γG. It thus appears that anti-protein A reactivity is confined to γG-1, γG-2 and γG-4 molecules.
Immunoglobulin Production in Synchronized Cultures of Human Hematopoietic Cell Lines
Authors: Takahashi M. , Yagi Y. , Moore G. E. , Pressman D.
Published: December 2022

The production of immunoglobulin in the various metabolic stages of the generation cycle of cultured human lymphocytoid cells was investigated by the use of synchronized cultures. The cells were synchronized at the boundary of the G1 and S periods by treatment with excess thymidine and the cells harvested at different time intervals after the release from the thymidine treatment were examined for their immunoglobulin content. The content of immunoglobulin was determined for individual cells by treating cell smears with 125I-labeled anti-immunoglobulin antibody, submitting them to radioautography and counting the silver grains on individual cells. Cell smears were also stained with fluorescein-conjugates of antiimmunoglobulin antibodies.

Although the immunoglobulin content of individual cells differed greatly even in the same harvest of a synchronized culture, there was a general change during the course of the generation cycle. The immunoglobulin level increased rapidly after the release of the thymidine blockage and reached its peak in the early portion of the S period. It then decreased gradually through the late S and G2 periods and reached its lowest level in the mitotic period. The second phase of increase which was observed after the mitotic burst indicated the synthesis of immunoglobulin by the cells of the second generation in the G1 and early S periods.

Further studies on the secretion of the immunoglobulin are required to obtain an overall pattern of the immunoglobulin synthesis.
Immunologic Cross-Reactivity of Escherichia Coli B Envelope Glycoproteins with Some Animal and Plant Cell Membrane Proteins
Authors: Okuda Shinichi , Weinbaum George
Published: December 2022

During the past few years a number of laboratories have been investigating the antigenic character of various components of the bacterial envelope. The goals are to develop vaccines against pathogenic organisms (1) and to understand more completely the structural organization of the cell surface (2). We recently reported that rabbit antiserum prepared against isolated envelopes of Escherichia coli B had immunologic reactivity with certain animal cell membrane proteins (3). Specifically, the anti-envelope serum reacted with a “structural protein” preparation from beef heart mitochondria and a phenol soluble protein fraction from the same type of mitochondria. The phenol solubilization technique has been shown to be extremely effective in isolating membrane specific proteins (3), solubilizing membranes such as erythrocyte ghosts which are normally quite resistant to most procedures (4) and preferentially solubilizing glycoproteins (5).
Studies of Lymphocyte Culture: Products of Sensitive Lymphocyte-Antigen Interaction
Authors: Spitler Lynn E. , Lawrence H. S.
Published: December 2022

The biologic activities of substances produced during stimulation of sensitized blood lymphocytes by specific antigen were studied with the following results.

1. A factor causing inhibition of migration of normal macrophages is produced, and, in the system used, the degree of inhibition is proportional to the degree of lymphocyte stimulation as measured by radioactive thymidine incorporation.

2. The activity of the inhibitory factor was increased in some experiments by increasing the dose of specific antigens.

3. Incubation of sensitive lymphocytes with antigen produces supernatants, which, when added to normal lymphocytes, cause them to respond with increased thymidine uptake. The resultant dose-response curve observed in normal cells exposed to such supernatant preparations parallels that observed when sensitive lymphocytes are cultured in the presence of antigen.
Serologic Classification of Chimpanzee Adenoviruses by Hemagglutination and Hemagglutination Inhibition
Authors: Hillis William D. , Goodman Rosanne
Published: December 2022

Twenty-two strains of viruses, each possessing adenovirus complement-fixing antigen and each derived from fecal specimens of chimpanzees, have been classified by their hemagglutinating properties into three major subgroups. Subgroup I is composed of eight strains which cause hemagglutination of rhesus or vervet monkey erythrocytes; subgroup II consists of 12 strains which agglutinate selected Sprague-Dawley rat erythrocytes in the presence of heterotypic immune serum to members of the Rosen subgroup III of human adenoviruses; and finally, chimpanzee subgroup III is composed of two strains which fail to agglutinate monkey, rat, guinea pig, or human type O erythrocytes.

HAI reciprocal tests suggest that all eight of the subgroup I strains are related to prototype C-1 chimpanzee adenovirus, as well as to human adenovirus type 16. Six strains of subgroup II are closely antigenically related to each other and to human adenovirus type 5 by HAI. Four additional strains are partially cross-reactive in HAI tests with human adenovirus type 2 and highly cross-reactive with one another. The remaining two strains of subgroup II apparently represent previously unreported serotypes without relationships to known adenoviruses or to each other, as demonstrable by HAI techniques. Evidence is presented that the 22 strains represent a total of at least seven distinct chimpanzee serotypes of adenovirus.
The Antigenic Subunits of Influenza Viruses
Authors: de St. Groth S. Fazekas
Published: December 2022

The breadth of the antibody response to antigenic subunits and whole virus vaccines was compared on sera against eight influenza A strains. The spectrum of cross-reactions for the two kinds of vaccine was statistically indistinguishable, both in antihemagglutinin and in neutralization tests. A strong negative correlation was found between the capacity of an antigen to give broad immunologic coverage and its proneness to being inhibited by heterologous sera. A model is proposed to account for the asymmetry of serologic crossing, and its theoretically and practical implications are discussed.
Further Evidence of the Role of γl Guinea Pig Antibodies in Mediating Passive Cutaneous Anaphylaxis (PCA)
Authors: Nussenzweig Victor , Benacerraf Baruj , Ovary Zoltan
Published: December 2022

It has been previously shown that passive anaphylactic reactions can be produced in the guinea pig and the mouse by an electrophoretically fast moving homologous immunoglobulin which is usually designated as γ1 globulin (1–4). In some other species (man, rabbit, dog, monkey and rat) anaphylactic reactions are mediated by homologous reaginic antibodies which have physicochemical properties clearly different from those of γ1 globulin (5). In man, these antibodies have been identified as belonging to the γE class of immunoglobulins (6). In mice both γ1 and reaginic antibodies have been shown to mediate passive cutaneous anaphylaxis (PCA) (7–9). It has recently been suggested that PCA in the guinea pig is not mediated by γ1 globulin but by a contaminant of the γ1 fraction (10). The object of this study was to investigate this possibility.
Anti-Thymus Cytotoxicity: Identification and Isolation of Rat Thymus-Specific Membrane Antigens and Purification of the Corresponding Antibodies
Authors: Bachvaroff R. , Galdiero F. , Grabar P.
Published: December 2022

Two rat thymus-specific antigens inducing cytotoxic hetero-antibodies are localized in the cell membrane, endoplasmic reticulum and outer mitochondrial membrane. One is also common to the cell-sap. The two antigens were purified from detergent-solubilized endoplasmic reticulum by gel filtration on Sephadex G-200 and preparative polyacrylamide disc electrophoresis until single homogeneous electrophoretic protein bands were obtained. Some properties of the unique membrane antigen suggest that it may belong to the group of the structural membrane proteins.

The firm binding of the antigens to the membranes permitted absorption and subsequent elution of cytotoxic antibodies from the whole immunoserum previously rendered thymus-specific by absorption with subcellular fractions from different organs. The eluted antibodies were exclusively IgG and retained their activity in vitro and in vivo.

The results are considered relevant to the study of the mechanism of the immuno-depressive action of thymus-specific cytotoxic antibodies.
Isolation and Characterization of Free λ-Chain of Immunoglobulin Produced by an Established Cell Line of Human Myeloma Cell Origin
Authors: Matsuoka Y. , Yagi Y. , Moore G. E. , Pressman D.
Published: December 2022

A λ-type light chain of immunoglobulin was isolated in an essentially pure state from the cells of an established cell line (No. 8226) derived from a myeloma patient S and compared with the λ-chain isolated from the spent culture medium. The cellular λ-chain was found to be essentially identical to the λ-chain secreted into the medium with respect to its physical, chemical and antigenic properties. Evidence was obtained that the λ-chains are present in the cell extract as a covalently-linked dimer although the purified preparation showed the presence of both covalent and noncovalent dimers. Apparently the covalent dimers were converted into noncovalent dimers by reduction during chromatography on CM-cellulose.

The cells also contained a substance that is related antigenically to the λ-chain of this cell line, but is smaller in molecular size than the completed λ-chain. This substance may represent a portion of the variable portion because it possessed only part of the idiotypic determinants and no determinants common to other λ-chains.
Impaired Lymphocyte Transformation in Ataxia-Telangiectasia in Part Due to a Plasma Inhibitory Factor
Authors: McFarlin Dale E. , Oppenheim Joost J.
Published: December 2022

Studies of lymphocyte transformation in ataxia telangiectasia (A-T) revealed reduced responses to PHA in 14 of 16 cases, to PWM in eight of 12 cases, and to SLO in seven of 10 cases, when 20% autologous plasma was used in the culture media.

It was possible to improve the responses of lymphocytes from 11 of 14 patients with subnormal responses to PHA and from six of eight having reduced responses to PWM by either increasing the concentration of the respective stimulant or by culturing in 20% homologous plasma. When both of these were used simultaneously their beneficial effects were additive. Tenfold increases in the optimal concentrations of PHA and PWM, which were somewhat toxic to normal lymphocytes, resulted in increased tritiated thymidine incorporation by A-T lymphocytes. No significant improvement in the reduced responses to SLO was obtained by either culturing in homologous plasma or by increasing the concentration of this stimulant.

Plasma from some A-T patients inhibited the response of normal control lymphocytes to PHA and to a lesser extent to PWM. The relationship of plasma inhibition to the concentration of these stimulants was consistent with a competitive mechanism of action.

The sera which inhibited the response of normal cells to PHA and PWM had elevated levels of α2 globulin on paper electrophoresis, which raises the possibility that the inhibitory factor may have this electrophoretic mobility.

Although the inhibitory factor accounted in part for the reduced responses to PHA and PWM in approximately 57% of the cases, it did not provide an explanation for the reduced responses in the remaining 25% of the A-T cases with subnormal responses to these stimulants, nor was it shown to contribute to the reduced responses to SLO.
Passive Transfer of Experimental Allergic Encephalomyelitis: Mechanisms of Suppression
Authors: Falk George A. , Kies Marian W. , Alvord Elisworth C.
Published: December 2022

Passive (adoptive) transfer of experimental allergic encephalomyelitis (EAE) with lymph node cells has been used as a method of analysis of mechanisms of disease induction and protection. Pretreatment of cell donors with CNS tissue prior to active challenge yielded lymph node cells which were unable to induce EAE in normal recipients. Similar pretreatment of recipients with CNS before they were given cells from actively sensitized donors rendered them unreactive to an otherwise highly encephalitogenic cell inoculum. Interference with the formation of sensitized cells is thus only one part of and not the whole explanation for the phenomenon of protection. Mycobacteria-induced protection against EAE similarly created an internal environment in the recipients which did not permit passive transfer of EAE. In each case, protection against EAE was accompanied by inhibition of the delayed dermal reaction to the CNS encephalitogen, myelin basic protein (BP).

The site of interference with delayed hypersensitivity, which occurs at some point beyond the initial sensitization, was not defined by the present study. The protected state, which in most animals was not accompanied by detectable antibodies to the encephalitogen, could not be transferred passively to other animals with either serum or lymphoid cells from protected animals.
Reactivity of Polymer (19S) and Monomer (7S) Forms of γM with Anti-γM Serum
Authors: Gleich Gerald J. , Loegering David A.
Published: December 2022

An anti-γM serum which distinguishes between 19SγM and a low-molecularweight γM from human serum (γMs-N) was prepared by immunizing a goat with normal human γM in Freund's complete adjuvant; it was absorbed with γG and hypogammaglobulinemic serum and was specific for γM. The reactivity of this antiserum was examined with 19SγM, with the γM monomer produced by reduction and alkylation (γMs-RA), and with the γM monomer prepared by reduction alone (γMs-R). In agar double diffusion the 19SγM band spurred over the bands formed by γMs-RA and γMs-R whereas the bands formed by γM monomers fused. Mixtures of 19SγM and γMs-RA formed concentric zones of precipitation in radial immunodiffusion and produced two bands in agar double diffusion. By quantitative precipitin analysis, with 19SγM the antiserum contained 30 µg Ab N/0.1 ml serum and with γMs-RA, 19 µg Ab N/0.1 ml. After absorption at equivalence with γMs-RA the antiserum still precipitated with 19SγM and Fc-γM but did not precipitate with Fab-γM. The precipitin reaction between 19SγM and the anti-γM serum absorbed with γMs-RA was inhibited by three γM subunits (γMs-RA, γMs-R, and γMs-N). Thus, the antiserum is reactive with an antigenic determinant on the Fc portion of the µ chain present on both 19SγM and the γM subunits. The difference in the reactivity of the antiserum with 19SγM and the γM subunits may be related to the number of specific antigenic determinants on these molecules.