bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigen-Specific T Cell-Mediated Suppression
Authors: Germain Ronald N. , Theze Jacques , Waltenbaugh Carl , Dorf Martin E. , Benacerraf Baruj
Published: December 2022

The induction of new suppressor T cells (Ts2) by suppressive extracts (TsF)from L-glutamic acid50-L-tyrosine50 (GT) nonresponder mice was examined. Incubation of normal spleen cells with allogeneic GT-TsF for 2 days in vitro led to the generation of Ts2 cells able to suppress subsequent responses to the immunogen GT-methylated bovine serum albumin (GT-MBSA) in vivo. This induction occurred efficiently when TsF donor and target cells differed at all of H-2, including the I-J subregion. B10.BR (H-2k) GT-TsF, adsorbed on, then acid eluted from GT-Sepharose and anti-I-Jk [B10.A (3R) anti-B10.A (5R)]-Sepharose in a sequential fashion could induce BALB/c (H-2d) spleen cells to become Ts2 only if nanogram quantities of GT were added to the purified GT-TsF. This indicates a requirement for a molecule or molecular complex possessing both I-J determinants and antigen (GT)-binding specificity, together with GT itself, for Ts2 induction. The induced Ts2 are I-J+, since their function can be eliminated by treatment with anti-I-Jk plus C. These I-J determinants are coded for by the precursor of the Ts2 and do not represent passively adsorbed, I-J coded TsF, since anti-I-Jk antiserum [(3R × DBA/2)F1 anti-5R] which cannot recognize the BALB/c (I-Jd) TsF used for induction still eliminates the activity of induced A/J (I-Jk) Ts2. These data provide further evidence for and information about the minimum of two T cells involved in antigen-specific suppressor T cell systems.
Antigen-Specific T Cell-Mediated Suppression
Authors: Germain Ronald N. , Benacerraf Baruj
Published: December 2022

Previous studies demonstrated that nonresponder (DBA/1)-derived suppressor factor specific for the random copolymer L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT-TsF) failed to suppress responder strain mice when administred at the same time as antigen (GAT) challenge. The present study demonstrates that if responder (BALB/c or A/J) spleen cells are preincubated with nonresponder GAT-TsF for 2 days in vitro, suppressor T cells (Ts2) are induced that can suppress syngeneic GAT plaque-forming cell responses in vivo or in vitro. This finding is in accord with Ts2 induction in nonresponder mice with the same GAT-TsF. Further, administration of nonresponder GAT-TsF to responder mice 7 days before in vivo antigen challenge suppressed subsequent GAT-PFC responses in these mice. Finally, the Ts2 induced in the same strain of mice (BALB/c) by GAT-TsF and by L-glutamic acid50-L-tyrosine50 (GT)-specific TsF were shown to be able to suppress only the response to the inducing antigen, demonstrating the fine specificity of these Ts2. The implications of this demonstration of nonresponder TsF activity in responder mice for an antigen under strict Ir gene control are discussed.
Sharing of Ia Antigens Between Species
Authors: Shinohara Nobukata , Lunney Joan K. , Sachs David H.
Published: December 2022

A mouse alloantiserum B10.D2 anti-B10.BR (H-2d anti-H-2k) cross-reacted with rat lymphocyte surface glycoproteins with characteristics of Ia antigens. Sequential precipitation analysis on solubilized radiolabeled LEW rat lymphocyte antigens with this cross-reactive mouse alloantiserum and the rat alloantiserum BN anti-LEW (Ag-B3 anti-Ag-B1) revealed that the Ia-like antigen detected by the mouse alloantiserum also reacted with the rat anti-Ia antibodies. It was further shown that the rat alloantiserum also detected another set of Ia-like antigens that did not cross-react with the mouse alloantibody. Precipitation analysis with congenic rat strains confirmed that all Ia-like antigens precipitated by the rat alloantibody were encoded by Ag-B linked genes. Thus the shared Ia-like antigen must also be the product of Ag-B-linked gene(s) or be physically associated with such products. In addition, molecules bearing shared antigenic determinants were separable from at least some of the Ia-like antigens detected by the rat alloantiserum, possibly suggesting the existence of more than one sublocus coding for Ia antigens within the rat MHC.
Physicochemical Separation of Two Distinct Fc Receptors on Murine Macrophage-Like Cell Lines
Authors: Anderson Clark L. , Grey Howard M.
Published: December 2022

By using radiolabeled myeloma proteins specific for each of two distinct Fc receptor activities on macrophage-like murine cell lines (“aggregated IgG FcR” activity and “monomer IgG2a FcR” activity), we have been able to detect solubilized FcR of both types in detergent lysates of these cells. The two solubilized Fc receptors can be distinguished and physically separated from one another by means of affinity chromatography and by sucrose gradient centrifugation. They also differ in their sensitivity to phospholipase C. The detergent solubilized monomer IgG2a FcR can be removed from solution with a Sepharose IgG2a column, has an S value of 4 to 5 and is resistant to phospholipase C. The solubilized aggregated IgG FcR does not bind to insolubilized IgG2a, has an S value of greater than 19, and is sensitive to phospholipase C treatment. We conclude from these studies that two different cell membrane macromolecules are responsible for the Fc receptor activities observed in macrophage-like cell lines.
The Potential of Bursa-Immigrated Hematopoietic Precursor Cells to Differentiate to Functional B and T Cells
Authors: Weber W. T. , Alexander J. E.
Published: December 2022

The potential of hematopoietic precursor cells, recently immigrated into the 13- and 14-day-old embryonic bursa, to migrate to the thymus and to differentiate to functional T cells was investigated. Chromosomally marked cell populations obtained from 13- and 14-day-old embryonic bursas were transferred i.v. to 780 R γ-irradiated chick embryos of equivalent age. When appropriate chimeras were examined at 4 to 12 weeks after cell transfer, donor cells were found to proliferate primarily in the bursa. Significant donor cell influx into the thymus was not detected. In correlation with these findings, Con A- and PHA-responsive T cells in thymus and spleen cell cultures of recipients remained of host origin whereas the number of anti-CIg responsive B cells of donor type increased gradually in the spleens of recipients. An initial lag period preceded the accumulation of functional donor B cells in the spleens of recipients, despite the predominant presence of dividing donor cells in the bursa. This suggests that the transferred bursal cell population required substantially longer to mature and emigrate from the bursa as functional B cells than the host cell population remaining in the irradiated bursas at time of cell transfer. The failure to detect significant influx of donor cells into the thymus and their failure to differentiate to functional T cells suggest that the recently bursa-immigrated hematopoietic stem cells of 13- and 14-day-old embryos may not be pluripotential cells, but rather cells already committed to the B cell line of differentiation.
Evidence That Bovine Conglutinin Reacts with an Early Product of C3b Degradation, and an Improved Conglutination Assay
Authors: Linscott William D. , Ranken Raymond , Triglia Richard P.
Published: December 2022

When EAC43b were treated with heated serum in EDTA, reactivity with bovine conglutinin appeared rapidly, even at 0°C, and almost simultaneously with the loss of C3b rosetting capacity. At the time conglutinability first appeared, there was no detectable decrease in I-A or hemolytic C3 activity, and no detectable C3 antigen release from the cells. With prolonged exposure to heated serum in EDTA, I-A (immune adherence) and hemolytic C3 activity were lost. If this exposure was at 37°C, C3 antigen became strongly detectable in the supernatant fluid, and eventually conglutinability was markedly reduced or lost, whereas C3d rosettes were unaffected. We suggest that bovine conglutinin reacts with some early product of C3b degradation, rather than with C3d, and propose that this intermediate be designated C3k.

We have developed a semi-quantitative assay for bovine conglutinin, utilizing a Coulter Counter to register the decrease in total particles due to red cell aggregation. By using this method, we have detected conglutination with mouse complement (C) as well as with that from man and the guinea pig.
Human Lymphocyte Responses Are Enhanced by Culture at 40°C
Authors: Smith J. Bruce , Knowlton Regina P. , Agarwal Shyam S.
Published: December 2022

In vitro responses of human peripheral blood lymphocytes (PBL) were found to be markedly enhanced by culture at 40°C rather than at the conventional temperature of 37°C. We studied proliferative responses of lymphocytes by activation by phytohemagglutinin (PHA) concanavalin A (Con A), pokeweed mitogen (PWM), and allogeneic lymphocytes in mixed lymphocyte culture (MLC) and found enhancement of DNA synthesis at the higher temperature. Cytotoxic T cell responses to allogeneic cells were also enhanced when MLC was done at 40°C. These enhanced immune responses appear to be due in part to increased numbers of participating cells. If in vitro lymphocyte responses correlate with in vivo responses, then fever associated with infection or tumor may be beneficial whereas that associated with autoimmune disorders may have a detrimental effect.
Target Antigens for H-2-Restricted Vesicular Stomatitis Virus-Specific Cytotoxic T Cells
Authors: Zinkernagel R. M. , Althage A. , Holland J.
Published: December 2022

Cytotoxic thymus-derived lymphocytes from mice infected with vesicular stomatitis virus (VSV) are H-2 restricted and virus specific for the Indiana and New Jersey strains of VSV. VSV-Indiana-immune T cells can lyse target cells infected with the temperature sensitive (ts) mutant ts 045 about 30 times better when target cell infection occurs at the permissive rather than the nonpermissive temperature. Since this mutant fails to express the glycoprotein at the cell surface when grown at the nonpermissive temperature, our results support the view that the viral glycoprotein is involved in defining the major target antigen for VSV-specific T cells. However, the tl 17 mutant that expresses a mutant glycoprotein at the cell surface was lysed, suggesting that the expressed mutated glycoprotein can cross-react with Indiana wild-type glycoprotein. Targets infected at the nonpermissive temperature with VSV ts G31 (mutant in the matrix protein) are still susceptible to T cell-mediated lysis but at a lower level of sensitivity. These results are compatible with the interpretation that for VSV-specific T cell lysis of infected target cells, the viral glycoprotein is a major target antigen and must be expressed, and that the matrix protein plays a lesser role, probably by indirectly influencing glycoprotein configuration at the cell surface.
Structural Studies on Protein Products of Murine Chromosome 17
Authors: Cook Richard G. , Vitetta Ellen S. , Uhr Jonathan W. , Klein Jan , Wilde Charles E. , Capra J. Donald
Published: December 2022

Twenty-one of the amino terminal twenty-seven amino acid residues have been assigned in the murine H-2Kq molecule and several additional assignments and corrections have been made in both the H-2Kk and H-2Kb molecules. The study shows that 1) H-2Kk and H-Kq, which are serologically related, have an identical amino acid sequence for all twenty positions that can be compared, 2) the H-2Kk and H-2Kq molecules differ from H-2Kb at only two positions-9 and 22, and 3) the additional sequence data on the H-2Kk, -Kb, and -Kq molecules increases the already striking homology between murine and human transplantation antigens.
Antigen-Specific Regulation of Myeloma Cell Differentiation in Vivo by Carrier-Specific T Cell Factors and Macrophages
Authors: Rohrer James W. , Lynch Richard G.
Published: December 2022

Previous studies demonstrated that: i) the TNP-binding myeloma MOPC-315 differentiated during in vivo growth in diffusion chambers (DC) implanted i.p. into normal BALB/c mice, and ii) the myeloma cell differentiation was regulatable by carrier-specific presentation of TNP to MOPC-315 cells in carrier-primed mice. In those studies, promotion and suppression of MOPC-315 differentiation occurred in the presence of carrier-specific helper and suppressor activities, respectively.

In the present studies, we demonstrate that carrierspecific regulation of MOPC-315 differentiation can be adoptively transferred to normal mice with carrierprimed T lymphocytes. In addition, the induced regulation of MOPC-315 differentiation is abrogated when macrophages are not present with MOPC-315 cells in the DC. These studies establish the immunologic basis of myeloma cell regulation and suggest that soluble, carrierspecific helper and suppressor factors of T cell origin regulate MOPC-315 differentiation directly or in collaboration with macrophages.
Antigen-Induced Increase in Migration of Large Cortical Thymocytes (Regulatory Cells?) to the Marginal Zone and Red Pulp of the Spleen
Authors: Durkin Helen G. , Carboni Joan M. , Waksman Byron H.
Published: December 2022

Antigen-dependent and antigen-independent thymocyte migrations are described. Rats injected i.v. with a large dose of ovalbumin (100 mg) showed a 3-fold increase in the numbers of large, low density cells in thymus 2 days later. When subpopulations of different density were labeled with 3H-adenosine and transferred to normal recipients, they migrated preferentially to spleen, unlike cells from saline-injected donors, which equilibrated among all lymphoid organs. Data from autoradiographs and dissection of spleens showed that the increased radioactivity in spleen resulted from a 3-fold increase in the number of large cells, predominantly in the marginal zone and, secondarily, in the red pulp. This increased migration to marginal zone with antigen was superimposed upon a normal pathway taken by large, low density thymocytes to this site. When adrenalectomized rats, similarly treated, received intrathymic 125IUdR, radioactivity in spleens of the ovalbumin group after 24 hr was 2- to 3-fold greater than controls and, again, associated only with marginal zone-red pulp. When only cortical thymocytes were labeled by topical application of 125IUdR to the thymus capsule, the same enhanced localization in spleen was seen at 24 hr. Migration of small, dense, medullary thymocytes to the periarteriolar lymphoid sheath (classical thymus-dependent area) was unaffected by antigen.

Only large, low density thymocytes migrated to Peyer's patches. This migration was unaffected by antigen. Only dense thymocytes localized in the thymus. However, if thymocytes were transferred from antigentreated donors, no subpopulation homed to thymus.
Positive and Negative Allogeneic Effects Mediated by MLR-Primed Lymphocytes: Quantitation by Limiting Dilution Analysis
Authors: Corley Ronald B. , Kindred Berenice , Lefkovits Ivan
Published: December 2022

Lymphocyte populations primed in mixed lymphocyte cultures have been used to determine if the allogeneic effect is mediated by alloreactive T lymphocytes. A microculture system with excess B cells, sheep erythrocytes, and limiting numbers of allogeneic T cells was used to compare the frequencies of allogeneic “helper” cells in primed and unprimed populations. A positive allogeneic effect, i.e., the production of anti-sheep erythrocyte antibodies, could be detected when low numbers of primed T cells were added to the cultures. After priming to the H-2 antigens of the B cells used in the cultures, the frequency of allogeneic helper cells was increased at least 10-fold to approximately one in every 30 cells. Priming to different H-2 antigens did not produce this effect. Therefore, the allogeneic effect can clearly be mediated by cells that specifically recognized and respond to alloantigens.

As the number of primed T cells increased, the number of responding microcultures decreased. Lymphocytes mediating the suppressive effect were clearly different from those mediating positive effects with respect to their susceptibility to irradiation and mitomycin-C treatment. The frequency of the set of cells mediating the negative effect was estimated, but varied from experiment to experiment. That the helper activity could be detected only under conditions of limiting T cells stresses the importance of limiting dilution analysis in the functional assessment of mixed populations of cells.
Molecular Characterization of the Ss and Slp (C4) Proteins of the Mouse H-2 Complex: Subunit Composition, Chain Size Polymorphism, and an Intracellular (Pro-Ss) Precursor
Authors: Roos Marleen H. , Atkinson John P. , Shreffler Donald C.
Published: December 2022

Adherent peritoneal cells, presumably macrophages, from mice of a number of congenic strains when cultured in vitro in medium containing 14C-labeled amino acids were found to synthesize the Ss protein (mouse C4), as revealed by immunoprecipitation with anti-Ss serum from 14C-labeled, 24-hr culture supernatants. Strain B10.WR7, which expresses the Slp allotype of the Ss protein, was shown by the same method also to synthesize the Slp antigen. SDS-solubilization and 2-ME-reduction of the B10.WR7 immunoprecipitates followed by separation on SDS polyacrylamide slab gels and autoradiography revealed three major polypeptide bands in Slp-positive Ss molecules and three major bands in Slp-negative Ss molecules with m.w. comparable to those of the α-, β-, and γ-chains of human C4. The Slp-negative Ss polypeptides of strain B10.WR7 had apparent m.w. of 98,000, 75,000, and 33,500 whereas the Slp-positive Ss molecules of B10.WR7 yielded three polypeptides with apparent m.w. of 105,500, 75,000, and 32,000. Immunoprecipitation analyses of normal, unlabeled B10.WR7 serum or plasma in SDS slab gels followed by staining with Coomassie Blue revealed similar differences in the m.w. of the α- and γ-polypeptides of the Slp-negative versus the Slp-positive Ss proteins. Such analyses of sera and plasmas of mice from a backcross linkage test and from a series of intra-H-2 recombinants showed clearly that the m.w. differences in α- and γ-chains are controlled by the S region of the H-2 complex. In addition, the Slp-negative Ss protein in strains other than B10.WR7 had an Ssα-chain of 103,000 daltons, indicating the existence of still further Ss polymorphism also controlled by the S region. An apparent intracellular precursor polypeptide of the Ss molecule with a m.w. of approximately 185,000 (comparable to the guinea pig Pro-C4 described recently) has also been demonstrated in lysates of cultured peritoneal cells. These results further confirm the structural homologies of the mouse Ss-Slp and human and guinea pig C4 proteins. They also indicate that Ss and Slp-antisera define two similar, but structurally distinct subclasses of mouse C4 that are apparently controlled by two discrete structural genes in the S regions of Slppositive strains. It is postulated that each of these structural genes codes for a single polypeptide precursor for the native Slp-positive and Slp-negative Ss molecules, and that the observed differences in the m.w. of the α- and γ-chains of these two Ss subclasses reflect a mutational shift in the site of the proteolytic cleavage step that releases the chains.
Surface Immunoglobulin on Frog Lymphocytes
Authors: Mattes M. Jules , Steiner Lisa A.
Published: December 2022

The presence of surface immunoglobulin (Ig) on Rana catesbeiana lymphocytes was evaluated by immunofluorescence. With all antisera to frog low molecular weight (LMW) Ig and with certain antisera to high molecular weight (HMW) Ig, two populations of lymphocytes, one with and one without surface Ig, were identified. Both populations were present in all lymphoid organs except for the thymus, which had less than 5% Ig-bearing cells. Nonlymphoid cells were not stained. This distribution suggests that these populations are homologous to mammalian and avian B and T lymphocytes. Most cells with surface Ig bear HMW Ig, but a minority bear LMW Ig, and of these a large fraction is distinct from typical lymphocytes morphologically. Unexpectedly, other antisera to HMW Ig, but no control antisera, stained thymocytes, all lymphocytes, and also all other blood cells. This reaction was abolished by absorption with several preparations of HMW Ig purified in different ways. Absorption with frog erythrocytes also abolished the staining of thymocytes and nonlymphoid blood cells, but some lymphocytes continued to be brightly stained; absorption with a non-Ig component of frog plasma had the same effect. Evidently, similar determinants are present on HMW Ig, on a non-Ig plasma component, and on thymocytes and all blood cells. The presence of antibodies to such cross-reactive determinants in certain anti-Ig antisera may lead to the mistaken identification of Ig on cell surfaces.
Further Studies on the Tumor-Specific Suppressor Cells Induced by Ultraviolet Radiation
Authors: Fisher Michael S. , Kripke Margaret L.
Published: December 2022

Recent studies have suggested that the immunologic unresponsiveness of UV-irradiated mice against UV-induced fibrosarcomas might be due to the presence of suppressor lymphoid cells. In these experiments, we present additional evidence that suppressor lymphoid cells are present in lymph nodes and spleens of UV-irradiated mice and demonstrate that these cells are enriched after incubation on nylon wool columns, that they are inactive after treatment with anti-ϑ serum and complement, and that they are effective for at least 7 weeks after transfer to lethally x-irradiated mice. Splenectomy of UV-irradiated mice before tumor challenge did not restore their anti-tumor reactivity. The UV-induced suppressor cells appear to be specific for syngeneic UV-induced tumors, since they did not suppress the rejection of an allogeneic UV-induced tumor or two chemically induced syngeneic tumors.
Nature of the Antigenic Complex Recognized by T Lymphocytes
Authors: Thomas David W. , Shevach Ethan M.
Published: December 2022

In the present study we examined the effects of antisera directed against guinea pig Ia antigens on the ability of TNP-conjugated macrophages to stimulate TNP-specific T lymphocyte proliferation. Treatment of macrophages with anti-Ia sera for 1 hr before, 1 hr immediately after, or as late as 24 hr after TNP-modification resulted in a reduced ability to stimulate the TNP-specific T cell. The inhibition produced by anti-Ia sera was specific and did not result from interference with the ability of macrophages to process TNP-conjugated membrane antigens in a nonspecific manner. Brief treatment with anti-Ia serum did not result in inhibition of Ia-antigen synthesis nor could evidence of carry-over of anti-Ia antibody into the lymphocyte cultures be obtained. These results demonstrate that anti-Ia sera interfere with the development of a TNP-specific immunogen on the macrophage surface and strongly suggest that an association exists between TNP-modified membrane proteins and Ia antigens on the macrophage surface.
Enhancement by Interferon of Lymphocyte Cytotoxicity in Normal Individuals to Cells of Human Lymphoblastoid Lines
Authors: Skurkovich Semen V. , Skorikova Albina S. , Eremkina Elena I.
Published: December 2022

The cytotoxic activity of healthy donor lymphocytes toward cells of two human lymphoblastoid lines was studied by radioimmunoassay with 51Cr. Cells of the following lymphoblastoid lines were used as target cells: line L-101, derived from the peripheral blood of a patient with acute erythromyelosis, and line D-102, derived from the blood of a healthy donor. Lymphocytes of different donors revealed a certain cytotoxicity toward these target cells (from 0 to 34.4% for cells of L-101 line and from 3.1 to 37.2% for cells of D-102 line). Attempts to enhance the mentioned cytotoxic effect with interferon were made. Human leukocyte interferon was obtained by induction with Newcastle disease virus. The preparations of interferon were concentrated 10 to 20 times by ultrafiltration. As a control, preparations of “false interferon” were used, having been obtained and concentrated in the way analogous to interferon obtaining, except for viral induction. Preincubation of donor lymphocytes in interferon of different dilutions during 1 to 2 hr under 37°C led to enhancing their cytotoxic action toward lymphoblast target cells 1.5 to 30.6% higher in comparison with preincubation in the medium RPMI 1640 or in “false interferon.” A certain enhancing effect of “false interferon” preparations on the cytotoxicity of normal human lymphocytes toward lymphoblastoid target cells was observed, but in these cases the cytotoxicity value was significantly lower than that under the treatment with interferon. The probable role of interferon in immune reactions between lymphocytes and target cells is discussed.
Occurrence of Idiotypically Identical Antibodies in the Sera of Two Outbred Rabbits Hyperimmunized with Type II Pneumococcal Vaccine
Authors: Brandt Daniel Ch. , Jaton Jean-Claude
Published: December 2022

Thirty outbred New Zealand White rabbits were hyperimmunized i.v. with type II pneumococcal vaccine. Whole antisera were analyzed by isoelectric focusing (IEF) in the pH range of 5 to 9. Dissimilar IEF patterns were recorded for most of them with the exception of two rabbit antisera, 4422 and 311, which exhibited two antibody components with an indistinguishable IEF pattern. The antiserum 4422 was highly electrophoretically restricted, whereas antiserum 311 displayed several antibody components. Antibody components from antiserum 311 were isolated by affinity chromatography and the fraction exhibiting an IEF pattern indistinguishable from that of antibody 4422 was found to bind a nonasaccharide hapten derived from SII pneumococcal polysaccharide with the same affinity as antibody 4422. Both antibodies expressed a1/b4 allotypic specificities. Crossrecombinant molecules between antibodies 4422 and 311 exhibited the same affinity constant as that of parent antibody molecules, thus strongly suggesting that these have the same antigen complementarity-determining regions. Anti-idiotypic sera were prepared in guinea pigs against both antibodies. The reactions between 125I-labeled antibodies and their corresponding absorbed antiidiotypic serum were inhibited to the same extent by unlabeled antibody 4422 or 311, an observation that is consistent with the expression of full idiotypic crossreactivity between this pair. No inhibition was obtained with four other anti-SII antibodies of IEF patterns different from those of 4422 or 311. The guinea pig antiidiotypic sera were not antigen-binding site specific since the nonasaccharide hapten was not inhibitory in this system. Taken together, the results strongly suggest a high degree of structural similarity of two antibodies from outbred rabbits.
Identical VL Region Sequences of Two Antibodies from Two Outbred Rabbits Exhibiting Complete Idiotypic Cross-Reactivity and Probably the Same Antigen-Binding Site Fine Structure
Authors: Brandt Daniel Ch. , Jaton Jean-Claude
Published: December 2022

The amino acid sequences of the VL domains of two antibodies (4422 and 311) that exhibited the same isoelectric focusing (IEF) spectrotype, a1/b4 allotype, idiotype, and association constant for a nonasaccharide hapten have been determined. Two large fragments were obtained upon tryptic digestion of the citraconylated, fully reduced, and alkylated light chains owing to the presence of only two arginine residues per molecule of L chain. By homology with other rabbit k chain sequences, these peptides encompassed residues 1 to 63 and 64 to 214, respectively. The sequence of the N-terminal 60 residues was analyzed by automated Edman degradation with the whole L chain and by carboxypeptidases A + B digestions performed with the N-terminal 63-residue fragment. The citraconyl-peptide 64 to 214 was successfully degraded for 50 steps, which covered the sequence of the third hypervariable region. Both VL domains of antibodies 4422 and 311 were structurally identical with the possible exception of a Glu/Gln substitution at position 91. To our knowledge, this is the first report of two identical rabbit VK regions derived from antibodies expressing identical idiotypic determinants. In contrast, light chains from two other anti-SII antibodies, exhibiting IEF spectrotypes different from that of antibody 4422 (311), differed markedly both in their framework and first hypervariable regions from the leading sequence 4422. Taken together with the immunochemical data presented in the accompanying paper, these sequence data provide further strong support for the great similarity if not identity between antibodies 4422 and 311. Genetic implications of these results are discussed.
Presidential Address to the American Association of Immunologists, Delivered in Atlanta, Georgia, June 6, 1978
Authors: Austen K. Frank
Published: December 2022

The effector pathways recruited for host defense in both the nonimmune and the immune host are specific and generate an array of beneficial functions, but they can also be detrimental to the host if the response is misdirected or excessive. A maladjustment in function of these effector systems so as to contribute to pathologic states of hypersensitivity is not to be viewed as an excessive physiologic response but rather as an involuntary response to an insult extrinsic to the system and of such a nature as to violate the usual mechanisms for maintaining integrity of the host. My growing awareness of the physiologic rather than the pathologic potential of two such systems, namely the mast cell-eosinophil axis, which is situated at portals of possible host penetration by hostile elements, and the alternative complement pathway which distinguishes self from nonself by biochemical rather than immunochemical means, prompts me to emphasize their homeostatic functions.
Selective Sensitivity to Hydrocortisone of Regulatory Functions that Determine the Magnitude of the Antibody Response to Type III Pneumococcal Polysaccharide
Authors: Markham Richard B. , Stashak Philip W. , Prescott Benjamin , Amsbaugh Diana F. , Baker Phillip J.
Published: December 2022

Previous studies on the basis for the immunosuppressive potential of adrenal corticosteroids have stressed that the effects of these agents on immune functions depend on the animal species being considered, as well as the subpopulations of lymphocytes involved in the expression of immune functions examined. In the present work, we have evaluated the effect of a single dose of hydrocortisone on three different immunoregulatory functions that can influence the magnitude of an antibody response to Type III pneumococcal polysaccharide (SSS-III) in mice; these functions include suppressor, amplifier, and helper activity that are dependent upon the presence of distinct subpopulations of thymus-derived (T) cells. The results obtained show that a single injection of a relatively large dose of hydrocortisone, when given at the time of priming with carrier, eliminated all evidence of carrier-specific helper T cell activity; hydrocortisone was also found to eliminate a significant amount of helper T cell activity when given after such activity had been generated. But, under the same experimental conditions, suppressor and amplifier T cell activities were unaffected, even in this steroid-sensitive species. Such selective sensitivity may account for some of the immunosuppressive potency of steroids.
Guinea Pig Homologues of TL and QA-2 Antigens
Authors: Schwartz Benjamin D. , Cigen Ragnhild , Berggard Ingemar , Shevach Ethan M.
Published: December 2022

The H-2, thymus-leukemia (TL), and Qa-2 antigens of mice are encoded by closely linked genes on murine chromosome 17, and have structural similarity in that each antigen is borne on a ≈ 44,000 dalton molecule associated with β2 microglobulin (β2µ). The extensive homology of major histocompatibility complex (MHC) products that exists for the mouse and guinea pig suggested that a similar homology might exist for products of genetic regions closely linked to the MHC. By taking advantage of the selective association of β2µ with H-2, Qa-2, and TL antigens, and by using the technique of sequential immunoprecipitation, we demonstrated two previously undescribed guinea pig molecules reactive with anti-guinea pig β2µ. The first molecule was composed of a 36,000 dalton glycoprotein associated with β2µ and was found on guinea pig thymocytes, but not lymphocytes. The second molecule was composed of a 40,000 dalton glycoprotein associated with β2µ, and was found on both guinea pig thymocytes and lymphocytes. By structure, chemical composition, association with β2µ, and tissue distribution, the first molecule is an attractive candidate for the guinea pig homologue of TL antigen, whereas the second fits the criteria for the guinea pig homologue of Qa-2 antigen.
Chronic Permeability of the Central Nervous System to Mononuclear Cells in Experimental Allergic Encephalomyelitis in the Lewis Rat
Authors: Stohl William , Gonatas Nicholas K.
Published: December 2022

In order to assess whether experimental allergic encephalomyelitis (EAE), a putative animal model for multiple sclerosis (MS), is an ongoing chronic disorder, we have studied the permeability of spinal cords of Lewis rats with EAE to 3H-uridine- or 3H-thymidine-labeled lymphoid cells obtained from thymuses of naive donors or from draining lymph nodes of donors injected with guinea pig spinal cord + complete Freund's adjuvant (CFA), guinea pig myelin basic protein + CFA, or with CFA alone. During the acute clinical phase of EAE there is a high-level infiltration of 3H-thymidine- or 3H-uridinelabeled cells into the spinal cords. After clinical recovery from EAE up to 58 days post-inoculation, there is a lowlevel infiltration of 3H-thymidine-labeled cells into the spinal cords. A similar infiltration into the spinal cords by 3H-uridine-labeled cells was not detected. Donor cells from animals immunized with CFA alone showed similar levels of infiltration into the spinal cords of animals with EAE as donor cells from animals immunized with the encephalitogenic emulsion. Spinal cords from recipients immunized with CFA alone showed no increased permeability to labeled cells. Heat-killed labeled cells did not migrate into the spinal cords of animals with EAE. We conclude that a) EAE is a chronic disease and in this regard is a valid model for MS; and b) in the chronic phase of EAE, recently divided cells (3H-thymidine-labeled cells) show higher levels of migration into the target tissue than 3H-uridine-labeled cells.
Serologic Analysis of Tumor-Associated Surface Antigens on a Moloney Leukemia Virus-Induced Lymphoma, MBL-2
Authors: Ng Ah-Kau , McIntire K. Robert , Holden Howard T. , Herberman Ronald B.
Published: December 2022

Tumor-associated surface antigens of a Moloney virus-induced lymphoma, MBL-2, were studied by a 51Cr release, humoral complement-dependent cytotoxicity assay. A syngeneic antiserum obtained from C57BL/6 (B6) animals 40 to 60 days after infection with Moloney sarcoma virus (MSV), and a xenogeneic antiserum prepared by immunizing rabbits with the MBL-2 cells were used in this study. The specificities of the MSV serum and the rabbit anti-MBL-2 serum were examined by the direct cytotoxicity assay with various target cells and by the cytotoxicity inhibition assay with MBL-2 as target cells. B6 leukemia cells, induced by Friend virus (FBL-3), Moloney virus (MBL-2), Rauscher virus (RBL-5), and benzpyrene [EL-4(G+)] could inhibit both sera, and a Gross virus-induced leukemia (E♂G2) was negative. BALB/c leukemias induced by Moloney virus (YC8 and LSTRA) and Rauscher virus (MCDV-12) as well as an A/SN strain leukemia induced by Moloney virus (YAC) also gave positive results whereas a radiation-induced BALB/c leukemia (RL♂1) was negative.

Similar results were obtained with the cytotoxicity inhibition assay when either soluble 3 M KCl extracts or whole cells were tested. Cell surface antigens extracted from MBL-2, RBL-5, and El-4(G+) were positive whereas extracts from E♂G2, normal spleen cells, and P815 (a mastocytoma of DBA/2 mice) were negative.

Despite the identical patterns of the specificity of the two antisera when whole tumor cells or crude extracts were tested, major differences in the specificity of the two antisera were noted when virus reagents were tested. Although the anti-MSV serum was inhibited by intact Moloney or Rauscher virus, the anti-MBL-2 serum was not inhibited by either intact or disrupted viruses. The anti-MSV and heterologous antisera recognized similar but not identical antigens with a distribution broader than the FMR (Friend-Moloney-Rauscher) specificity, but unrelated to Gross cell surface antigens (GCSA). The anti-MBL-2 may be detecting a Moloney virus-associated cell surface antigen that appears not to be a virion antigen.
Augmentation of Human Natural Cell-Mediated Cytotoxicity by a Soluble Factor
Authors: Koide Yukio , Takasugi Mitsuo
Published: December 2022

Natural cell-mediated cytotoxicity (NCMC) like immune T cell-mediated cytotoxicity and antibody production is regulated by a soluble factor released during coculture of lymphocytes with mitomycin C-treated lymphoblastoid cell lines. This N-cell-activating factor (NAF) enhances the activity of effector N cells and increases natural cytotoxicity. There appears to be no restriction for compatibility at the A and B locus of the major human histocompatibility complex in the production or activity of the factor. NAF was observed in the supernatant as soon as 2 days after initiation of mixed culture with a peak of production at 5 days. A soluble factor produced and released by T cells in response to stimulation by other cells acts by enhancing cytotoxicity of effector cells in NCMC, demonstrating a T-N cell cooperation.
Fc-IgM and Fc-IgG Receptors on Human Circulating B Lymphocytes
Authors: Pichler Werner J. , Broder Samuel
Published: December 2022

Peripheral blood B cells from normal humans were enriched with rosette techniques (mean 93.5% surface immunoglobulin positive). We studied these purified B cells for the expression of Fc-IgG and Fc-IgM receptors by using bovine red blood cells coated with IgG (EA-IgG) or IgM (EA-IgM). Between 81 and 94% (mean 88.67%) Fc-IgG and between 79 and 97% (mean 87.5%) Fc-IgM receptor positive cells were found by using this sensitive rosette technique.

The Fc-IgG receptor values had a tendency to decrease in culutre at 37°C. In contrast, cultivation at 37°C for 24, 48, and even 72 hr enhanced Fc-IgM receptor expression. Trypsinization (0.25%) removed Fc-IgM receptors, but not Fc-IgG receptors. However, after further cultivation of the trypsinized cells, Fc-IgM receptors were re-expressed on the same or even greater percentages of cells.

Fc-IgM receptors were also detected on monoclonal B cells (chronic lymphatic leukemia cells) expressing surface immunoglobulins with either δ or γ heavy chains exclusively. The high percentage of Fc-IgM positive cells in normal peripheral blood B cells, coupled with the observation that the Fc-IgM receptors are not confined to monoclonal B cells with particular classes of surface immunoglobulins, suggests that the Fc-IgM receptor may be present on essentially all B cells. Moreover, our results suggest that most human B cells expressed both Fc-IgM and Fc-IgG receptors simultaneously.
Amplification of the Activity of Human Leukocyte Inhibitory Factor (LIF) by the Generation of a Low Molecular Weight Inhibitor of PMN Leukocyte Chemotaxis
Authors: Goetzl Edward J. , Rocklin Ross E.
Published: December 2022

Leukocyte inhibitory factor (LIF), which was derived from human peripheral blood lymphocytes by stimulation with concanavalin A and partially purified by Sephadex G-100 gel filtration, inhibited the in vitro spontaneous migration and chemotaxis of human PMN leukocytes as assessed in a Boyden chamber micropore filter assay. The inhibitory activity was attributed to LIF, a principle defined in terms of its inhibition of PMN leukocyte migration from glass capillary tubes, since it was preferentially directed to PMN leukocytes as compared to mononuclear leukocytes, exhibited a size comparable to LIF by gel filtration, and was inactivated by diisopropyl fluorophosphate in parallel with LIF. Incubation of PMN leukocytes with LIF released additional inhibitory activity, distinct from LIF, which resembled the neutrophil-immobilizing factor (NIF) by virtue of its approximate m.w. of 4000 by filtration on Sephadex G-25, inactivation by trypsin digestion, and preferential noncytotoxic inhibition of spontaneous migration and chemotaxis of PMN leukocytes as compared to mononuclear leukocytes. Thus LIF inhibits PMN leukocyte migration both by a direct action on the cells and by an amplification pathway that is mediated by low m.w. chemotactic inhibitors similar to NIF.
Independent Analysis of T Helper and T Killer Cell Function in Young and Old NZB Mice
Authors: Falkoff Reuben Maverick , Scavulli John F. , Dutton Richard W.
Published: December 2022

With age, NZB mice lose their ability to develop a cytotoxic response after alloimmunization in vitro. This decline is shown to coincide with a diminution of T-helper cell activity as assessed by proliferation in mixed lymphocyte culture or in response to PHA. When cytotoxic T cell precursors are activated with the polyclonal activator Con A, there is no reduction in the number of cytotoxic effector T cells that develop. No autoreactive cytotoxic cells are seen in Con A-activated cultures. These findings are related to previous work on cell-mediated immunity in NZB and B/W mice.
Concanavalin A Increases Glyoxalase Enzyme Activities in Polymorphonuclear Leukocytes and Lymphocytes
Authors: Gillespie Elizabeth
Published: December 2022

Glyoxalase I converts methylglyoxal and glutathione to S-lactoylglutathione and glyoxalase II converts this compound to D-lactic acid, regenerating glutathione in the process. A recent study from my laboratory has provided evidence that S-lactoylglutathione modulates microtubule assembly in vitro whereas concanavalin A (Con A) has been shown to increase microtubule occurrence in polymorphonuclear leukocytes (PMN). The present report describes the dose-dependent activation by Con A of both glyoxalase I and II in PMN and lymphocytes. In nine experiments with PMN, Con A (100 µg/ml) increased glyoxalase I and II activities by 19 ± 8% and 12 ± 10% (mean ± S.D.). In 17 experiments with lymphocytes, activation of the two enzymes by 10 µg/ml Con A was 30 ± 14% and 28 ± 8%. Changes occurred after a 1-min incubation with Con A and persisted for at least 60 min. Since both enzyme activities are increased it is not clear if S-lactoylglutathione levels are increased or decreased but presumably they change. The present findings are compatible with the hypothesis that Con A increases microtubule occurrence in PMN by affecting the glyoxalase enzymes. They also represent a newly described early biochemical change caused by Con A in lymphocytes.
Lung Injury Produced by Immune Complexes of Varying Composition
Authors: Scherzer Herbert , Ward Peter A.
Published: December 2022

Immune complexes consisting of rabbit antibody to bovine serum albumin (BSA) have been made up at 1×, 3×, 6×, 8×, and 20× antigen equivalence. The complement fixing activity of these complexes is inversely proportional to the amount of antigen present in the complexes, and, as expected, solubility of the complexes progressively increases with increasing amounts of antigen. The ability of these complexes to induce acute pulmonary injury and inflammatory responses has been quantitatively assessed. Complexes preformed at antigen equivalence are the most damaging to lung, correlating with their complement fixing activity. When the antigen concentration in the complexes is increased 3 to 6 times beyond the point of equivalence, the phlogistic activity of the complexes drops off rapidly, as demonstrated by a sharp decline in the changes in vascular permeability, hemorrhage, and morphologic evidence of inflammation. These studies provide the first evidence that changing the physicochemical parameters of preformed immune complexes by simply altering the ratio of antigen to antibody can dramatically alter the phlogistic properties of immune complexes for pulmonary tissue.
Estimation of Antibodies Specific for Dextran
Authors: Matsuuchi Linda , Morrison Sherie L.
Published: December 2022

Methods are described for the isolation and characterization of picogram quantities of anti-dextran antibodies. 14C-dextrans produced by using the dextransucrases of Leuconostoc mesenteroides strains B1355 and B512 were used in a radioimmunoassay. The specificity of this assay was verified by using cell cytoplasmic lysates from mouse plasmacytomas, J558 (anti-α 1→3 dextran) and W3129 (anti-α 1→6 dextran). Dextran produced by strain B1355 and insolubilized with epichlorohydrin was used as an immunoabsorbent.
H-2 Linked Ir Gene Control of Antibody Responses to Porcine Insulin
Authors: Kapp Judith A. , Strayer David S.
Published: December 2022

Cell-mediated and humoral immune responses to heterologous insulins in mice are controlled by H-2-linked, dominant, immune response (Ir) genes. For example, mice bearing the H-2d haplotype develop T cell proliferative responses and produce antibody after injection with porcine insulin, whereas mice bearing other H-2 haplotypes do not. Data presented in this communication demonstrate that homozygous and heterozygous H-2d mice produce insulin-binding antibodies when immunized with porcine insulin or proinsulin. Some (H-2b,k,s) insulin-nonresponder mice produce insulin-binding antibodies after injection of proinsulin, whereas other insulin-nonresponder strains (H-2q) do not. All strains, except homozygous H-2q mice, produce antibodies specific for proinsulin, suggesting that the response to porcine proinsulin is also controlled by H-2-linked Ir genes. More importantly, F1 hybrids between insulin-nonresponder C57BL/10 (H-2b) and DBA/1 (H-2q) produce no insulin-binding antibodies when injected with proinsulin, despite the fact that proinsulin-binding antibodies are produced by these mice.
Specific Killing of Line-10 Cells in Vitro in the Presence of Normal Lymphoid Cells and Antibodies from Immune Rabbits or Tumor-Resistant Guinea Pigs
Authors: Brunda Michael J. , Minden Percy , Rodney Ferguson H.
Published: December 2022

Antibody-dependent cellular cytotoxicity (ADCC) was demonstrated against strain-2 guinea pig line-10 hepatocarcinoma cells by using syngeneic mineral oil-induced peritoneal exudate cells and rabbit antibodies to line-10 cells (anti-10). Cytotoxicity was dependent on the amount of anti-10 sera or IgG used. F(ab′)2 of rabbit anti-10 IgG was not active and cytotoxicity could be inhibited by addition to the assay of aggregated human IgG or of heat-killed Staphylococcus aureus containing protein A. Rabbit antibodies that were eluted from an immunoadsorbent to which antigens derived from line-10 cells were coupled mediated ADCC, whereas antibodies isolated similarly from an immunoadsorbent to which antigens drived from another syngeneic tumor, line-1, were coupled had little activity.

When 51 syngeneic sera from line-10 tumor-resistant guinea pigs were tested, a broad range of activity was observed and a large proportion of sera mediated ADCC. Immune sera were obtained from animals rendered tumor resistant by several different protocols, and it appeared that the method used to immunize animals influenced the level of ADCC observed. Sera from 44 normal guinea pigs had considerably less activity. Line-10-resistant guinea pig serum antibodies eluted from a line-10 immunoadsorbent mediated ADCC, whereas eluates of the same serum eluted from a line-1 immunoadsorbent had no activity. These data suggested that syngeneic line-10-specific antibodies in sera of tumor-resistant guinea pigs as well as rabbit anti-10 were capable of mediating ADCC in vitro.