bluuebunny/crossref_metadata_2025 huggingface dataset:
Antigen Induced Alterations in Splenic Prostaglandin and Cyclic Nucleotide Levels in NZB Mice
Authors: Webb D. R. , Nowowiejski I. , Dauphinée M. , Talal N.
Published: December 2022

Studies were undertaken in order to determine if NZB mice injected with sheep erythrocyte antigens would respond by showing elevated splenic prostaglandin and cyclic nucleotide levels similar to that observed in normal mice. The results show that young NZB mice can respond to sheep erythrocytes by yielding increased levels of splenic PGF2α and cAMP. However, because of increased basal levels of PGF2α and cAMP, the net increase observed is lower than that observed with normal mice. In old NZB mice exhibiting signs of disease (splenomegaly) and in which defects in immune competence are known to occur, the injection of sRBC results in no increase in splenic PGF2α levels and a decrease in cAMP levels. These animals also have greatly elevated basal levels of PGF2α, cAMP, and cGMP. It is concluded that the cellular immune defect in NZB mice is reflected by their faulty metabolic responses to sRBC. Also, the altered basal levels of PG and cyclic nucleotides may be related to the altered cellular immune competence. The latter conclusion is supported by the reduced capacity of spleen cells from young NZB mice to respond to PGE by increasing cAMP levels and by the lack of an effect of inhibitors of PG synthesis on the immune response to sRBC in both young and old NZB mouse spleen cell cultures.
Suppression of IgE Antibody Production in SJL Mice
Authors: Watanabe Naohiro , Kojima Somei , Shen Fung-Win , Ovary Zoltan
Published: December 2022

Under appropriate conditions of immunization combined with irradiation, SJL/J mice show a high and persistent anti-DNP IgE antibody response. Spleen cells transferred from normal untreated SJL mice suppress this response. Elimination of Ly-1+ cells, but not of Ly-2+ cells, abolished the capacity of spleen cells to suppress the IgE response.

Thus of the three T cell Ly subclasses presently identified, Ly-1, Ly-2,3, and Ly-1,2,3, the normal SJL spleen cell which suppresses the IgE response of irradiated-immunized SJL mice belongs to the Ly-1 set. It is not known whether this Ly-1 cell suppresses the IgE response directly or by helping another cell in the recipient.

The carrier-specific helper cell activity for IgE and probably IgG1 antibody response belongs to Ly-1 subclass in the SJL strain also.
Alternative Pathway of Complement: Nonenzymatic, Reversible Transition of Precursor to Active Properdin
Authors: Götze Otto , Medicus Rudolf G. , Müller-Eberhard Hans J.
Published: December 2022

A method for the purification of properdin in precursor state (pre-P) has been elaborated. Precursor properdin, in contrast to activated properdin (P̄), does not initiate the formation of a C3 convertase when added to properdin-depleted serum, rather it requires the presence of an activating substance such as zymosan for expression of its activity. Comparing the activities of pre-P and P̄ it was found that some P̄ preparations contained significant amounts of pre-P because they were fully active only in the presence of zymosan. This observation indicated that P̄ can partly revert to pre-P. Comparison by immunoelectrophoresis and sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed no charge or size differences between pre-P and P̄. Radiolabeled pre-P, when analyzed after its participation in the reactions of the pathway, displayed an unchanged subunit m.w. of 50,000. Taken together, these results strongly suggest that the transition of pre-P to P̄ is due to a partly reversible change in the conformation of the protein rather than the result of proteolytic cleavage.
Endothelial Proliferation in the Delayed Hypersensitivity Reaction: An Autoradiographic Study
Authors: Polverini Peter J. , Cotran Ramzi S. , Sholley Milton M.
Published: December 2022

The time-sequence of endothelial proliferation in the delayed hypersensitivity reaction was examined in the skin of guinea pigs. Animals were sensitized to tuberculin (PPD) and dinitrochlorobenzene (DNCB) and skin lesions were examined 12 hr to 7 days after challenge. Thirty minutes before sacrifice 3H-thymidine (3H-T) was injected i.p. and endothelial labeling was assessed in autoradiographs of 1.5-µ thick methacrylate sections. Endothelial labeling became manifest at 24 hr (PPD) and 12 hr (DNCB) after challenge, reaching a peak at 48 to 72 hr in PPD-sensitized animals and 36 to 48 hr in DNCB-sensitized animals. Peak labeling indices were 7.88 and 15.08%, respectively. Labeling indices for nonsensitized control groups were always less than 1%. Endothelial labeling was associated, in time of occurrence, distribution, and magnitude with mononuclear cell infiltration of the lesions. These findings thus document the occurrence of significant endothelial cell proliferation in delayed hypersensitivity responses and suggest that the events leading to the expression of hypersensitivity also result in endothelial replication.
The Mononuclear Cell in Human Blood Which Mediates Antibody-Dependent Cellular Cytotoxicity to Virusinfected Target Cells
Authors: Shore Steven L. , Melewicz F. Michael , Gordon David S.
Published: December 2022

Mononuclear cells (MC) from human blood were fractionated by a variety of physical and immunologic techniques, and the cellular subpopulations generated were assessed for their capacity to lyse herpes simplex virus (HSV)-infected target cells in the presence of IgG antibody to HSV. Latex phagocytosis and surface marker studies were performed in parallel in order to identify the major effector cells by their phagocytic properties and their possession of surface immunoglobulin and receptors for either sheep erythrocytes, C3, or the Fc fragment of IgG.

Cytotoxic effector cell activity was unaffected or slightly enhanced after the removal of plastic-adherent or carbonyl iron-adherent MC, indicating that the major effector cell is not a classical monocyte. Similar results were obtained after removal of more than 90% of the T cells by depletion of rosette-forming cells. Likewise, effector cell activity was generally unchanged when more than 95% of the B cells were removed by filtering MC on nylon wool columns. Effector cell function was also found to be normal in three patients with B cell-deficient X-linked agammaglobulinemia. These observations strongly suggest that the effector cells are not T cells or B cells.

A 4- to 5-fold enrichment in effector cells, however, was consistently found in a subpopulation, consisting of 5% of the unfractionated MC, that was dramatically enriched both for nonphagocytic cells with only Fc receptors (K cells) and for nonphagocytic cells with no detectable surface markers (null cells). Since, as is demonstrated in the accompanying report, effector surface Fc receptors play a critical role in the mediation of antibody-dependent cellular cytotoxicity directed at HSV-infected target cells, the major mononuclear effector cell in human blood is a K cell.
The Non-Specific Activation in the Complement Fixation Test of the Alcohol Soluble Antigens of the Tubercle Bacillus
Authors: Dienes L. , Scheff L. D.
Published: December 2022

It was found that the potency of certain preparations obtained from the alcohol extract of the tubercle bacillus as antigen in the complement fixation was increased 80 to 160 times by the addition of commercial lecithin. The potency of the original alcohol, ether, and watery extract was increased considerably less (2 to 10 times). Substances separated from the alcohol extract of the tubercle bacillus or cholesterin had no effect on the potency of the preparations. One millionth of a milligram of the specific substance (the dilution 1:400,000,000) in combination with lecithin was enough to cause as antigen a complete reaction in vitro. The significance and eventual consequences of these observations for the understanding of certain serological reactions are briefly discussed.
The Antigenic Properties of Pollen Fractions
Authors: Caulfeild A. H. W. , Cohen Cecile , Eadie George S.
Published: December 2022

Chemical methods have been given for the preparation from pollen of fractions which were dried and labelled as alcohol extract, albumin-proteose, proteose and glutenin. The percentage of total nitrogen was determined for these different fractions of ragweed and birch pollen. Both by means of death from anaphylactic shock and the uterine method of Dale it has been shown possible to sensitize guinea pigs with great ease and regularity to the albuminproteose, and proteose fractions of ragweed, timothy and birch pollen. The demonstration that glutenin or the alcohol extract can cause anaphylaxis is less complete owing to the less solvent character of these fractions. By means of the Dale technic marked sensitization was demonstrated with glutenin; the proof of sensitization with the alcohol extract being much less convincing. The demonstration of passive anaphylaxis from guinea pigs sensitized with albumin-proteose or proteose fractions to normal guinea pigs is regarded as complete; from human hayfever cases to normal guinea pigs, a slight sensitization resulted in two instances. Clinical tests by the scratch method demonstrated (a) that each case would show an individually selective specificity to the different fractions and (b) that positive results would occasionally be obtained when the use of whole pollen would give negative tests. Treatment by means of solutions made from ragweed fractions with a somewhat selected group of patients gave very satisfactory results. Although complete protection was not obtained in all, it was felt that the results were better than would have been obtained had but a single solution been used. It was entirely satisfactory in that all cases protected in previous years experienced as good or better results on their multiple solutions.
The Effect of D-Penicillamine on Mitogen-Induced Human Lymphocyte Proliferation: Synergistic Inhibition by D-Penicillamine and Copper Salts
Authors: Lipsky Peter E. , Ziff Morris
Published: December 2022

The effect of d-penicillamine on in vitro mitogen-triggered human peripheral blood lymphocyte DNA synthesis was examined. d-penicillamine caused a modest degree of inhibition of responsiveness. Since D-penicillamine avidly chelates copper, a possible additive effect of d-penicillamine and copper salts on mitogen-induced lymphocyte proliferation was investigated. The addition of various copper salts to D-penicillamine-containing cultures resulted in a marked augmentation in the degree of inhibition observed. Specificity for the synergy between D-penicillamine and copper was suggested by the failure of iron, zinc, or gold salts to augment the inhibition produced by D-penicillamine alone. However, a number of other thiols, but not disulfides, could substitute for D-penicillamine in inhibiting mitogen responsiveness in the presence of copper salts. Preincubation of peripheral blood mononuclear cells with d-penicillamine and CuSO4 resulted in diminished mitogen responsiveness on subsequent culture in the absence of the inhibitors. Data are presented to indicate that preincubation with d-penicillamine and CuSO4 directly affected the ability of the T lymphocytes to respond to mitogens but did not alter the capacity of the monocytes to act as accessory cells in this response.
Effect of Selective T Cell Priming on Anti-Sheep and Anti-Hapten Humoral Responses
Authors: Trizio Domenico , Cudkowicz Gustavo
Published: December 2022

Lymphocyte subpopulations responsible for immunologic memory developing within 4 days of priming with small numbers of sheep erythrocytes (2 × 106 SRBC, i.v.) were investigated by transfer of spleen cells into irradiated syngeneic mice, challenge with SRBC or SRBC-TNP conjugate, and enumeration of plaque-forming cells at the time of peak response. Cell suspensions were enriched with T and depleted of B lymphocytes, or vice versa, by filtration through nylon wool columns and separation of nonadherent from adherent cells. The characteristics of anti-SRBC or anti-TNP responses by primed unseparated cells (i.e., acceleration, augmentation, increased IgG:IgM ratio) were retained when T-rich spleen cells from primed mice were reconstituted with B-rich cells, irrespective of whether the latter were from primed or unprimed donors. Responses by unprimed T cells in combination with primed B cells resembled those elicited by unprimed unseparated spleen. Hence, low doses of antigen selectively activated the T lymphocyte subclass; helper cells became more effective after priming for cooperative interaction with B cells, especially with those committed to IgG production. Under conditions of limiting dilution for T cells, the helper units of primed mice (i.e., functional units of helpers and amplifiers) were restricted to interact with either IgM or IgG precursor B cells, but not with both, since direct and indirect plaque responses assorted independently of each other. The immediate precursors of primed helper cells were peripheralized T lymphocytes, in keeping with the persistence of helper activity for months after adult thymectomy. The incompetence of large numbers of transferred thymocytes to generate such helper cells was in sharp contrast with their prompt generation by small numbers of T-rich spleen cells.
Bronchopulmonary Macrophage Activation in the Pathogenesis of Hypersensitivity Pneumonitis
Authors: Stankus Richard P. , Cashner Frances M. , Salvaggio John E.
Published: December 2022

Repeated intratracheal (IT) inoculation of rabbits with a homogenized, saline suspension of Micropolyspora faeni produced bronchopulmonary (BP) histologic lesions resembling those of human hypersensitivity pneumonitis. With an in vitro phagocytic and bactericidal assay, an analysis of BP macrophages from M. faeni-injected rabbits demonstrated activation at both 2 and 4 weeks after the initiation of immunization. No BP macrophage activation was observed in immunized rabbits 6 weeks post-inoculation. BP macrophage activation was capable of recall after 6 weeks in M. faeni-sensitized animals that received a booster IT injection (2 mg) that did not activate “normal” alveolar wash cells. This recall of BP macrophage activation was accompanied by both a marked migration of mononuclear cells into the lung and positive delayed hypersensitivity skin reactions after intradermal injection of M. faeni antigen. Pulmonary histologic examination of sensitized, boosted rabbits suggested an enhanced cellular parenchymal infiltrate when compared with appropriate controls. The above observations confirm the occurrence of immunologically activated BP macrophages in rabbits inoculated with M. faeni via the respiratory tract route and suggest a correlation between macrophage activation and histopathology.
Effects of Restoring Lethally Irradiated Mice with Anti-Thy 1.2-Treated Bone Marrow: Graft-vs-Host, Host-vs-Graft, and Mitogen Reactivity
Authors: Norin Allen J. , Emeson Eugene E.
Published: December 2022

In vitro treatment of A/J mouse bone marrow with anti-Thy 1.2 serum and guinea pig complement (GPC) eliminated its ability to induce graft-vs-host (GVH) mortality in lethally irradiated C57BL/6J × A/J F1 (BAF1) mice. The anti-Thy 1.2 and GPC treatment of A/J marrow significantly reduced spleen cell activation by phytohemagglutinin (PHA) but not lipopolysaccharide (LPS) stimulation in A/J mice assayed 6 weeks after lethal irradiation and reconstitution with the treated marrow. However, the anti-Thy 1.2 treatment of A/J bone marrow did not impair the ability of the lethally irradiated, reconstituted, syngeneic mice to reject C57BL/6J skin grafts. We conclude that lymphocytes in bone marrow which are not susceptible to inactivation by anti-Thy 1.2 mediate allograft reactions and/or that radioresistant cells which persist in the recipient initiate rejection of allogeneic skin grafts.
Immunologic Characterization of Hairy Cell Leukemias in Continuous Culture
Authors: Saxon Andrew , Stevens Ronald H. , Quan Shirley G. , Golde David W.
Published: December 2022

The immunobiologic characteristics of three continuous cell lines established from hairy cell leukemia cells were investigated. All three cell lines continued to produce tartrate-resistant acid phosphatase, the enzymatic marker of hairy cells. Two of these cell lines were B lymphoid in nature. They carried Fc and C receptors, had surface and internal immunoglobulin, and did not form spontaneous sheep red blood cell rosettes. Experiments employing biosynthetic radiolabeling of immunoglobulin demonstrated distinctive immunoglobulin kinetics for each of these two hairy cell lines. One cell line remained quite similar to the original hairy cells from which it was derived whereas the other B lymphoid hairy cell line had undergone a switch in the immunoglobulin isotype produced. The third hairy cell leukemia line was shown to be of thymic derivation. These cells formed spontaneous sheep red blood cell rosettes and did not carry Fc or C receptors. The spontaneous sheep red blood cell rosette-forming cells contained tartrateresistant acid phosphatase. They did not possess surface or internal immunoglobulin and did not synthesize immunoglobulin in vitro. Hairy cell leukemia cells maintained in permanent cell culture retain their immunobiologic properties and offer the opportunity for indepth study of these unusual cells.
Complement-Dependent Stimulation of Normal Lymphocytes by Immune Complexes
Authors: Soderberg Lee S. F. , Coons Albert H.
Published: December 2022

Normal rabbit lymphocytes were stimulated to proliferate in vitro by antibody-antigen complexes. Stimulation was dependent upon C activity. Heat-inactivation or zymosan-treatment of the serum used in culture caused a 75 to 100% loss of responsiveness to the complexes. Serum-free culture or cultures with less than 1% serum supported only low levels of stimulation, but responsiveness reappeared proportionally with increased serum concentration. The low level dose-dependent responses seen in the absence of active C may have been due to C carried over with the cells or to stimulation independent of C. Aggregated rabbit γ-globulin tested over a broad dose range failed to stimulate normal lymphocytes more than minimally whether or not C was present. Stimulation with immune complexes was sustained by C4-deficient guinea pig serum, indicating participation of the alternative C pathway. Normal rabbit lymphocytes from peripheral blood, bone marrow, spleen, and lymph node proliferated in response to rabbit antibody-antigen complexes. Normal thymocytes were consistently unresponsive to the complexes.
Absence of Binding of MDP, a Synthetic Immunoadjuvant, to Anti-Peptidoglycan Antibodies
Authors: Audibert F. , Heymer B. , Gros C. , Schleifer K. H. , Seidl P. H. , Chedid L.
Published: December 2022

The binding of the synthetic immunoadjuvant N-acetyl-muramyl-l-alanyl-d-isoglutamine (MDP, for muramyl dipeptide) to human and rabbit sera containing peptidoglycan (PG) antibodies was investigated. Studies were performed by employing the corresponding 14C or 125I-labeled compounds in Farr-type binding and inhibition assays. Whereas MDP did not react with naturally occurring or experimentally induced PG-antibodies, the analog of MDP MDP-l-Lys-d-Ala did bind to hyperimmune rabbit anti-PG sera, but not to the human sera tested. These studies indicate that the radioimmunoassays employed basically are applicable for the selection of nonimmunogenic MDP analogs possessing immunoadjuvant activity.
Interactions Between Viruses and Lymphocytes
Authors: Doyle Michael V. , Oldstone Michael B. A.
Published: December 2022

Replicating virus was found in subpopulations of circulating blood mononuclear cells harvested from BALB/WEHI and SWR/J mice persistently infected with lymphocytic choriomeningitis virus (LCMV) either after intracerebral infection at birth or congenital in utero infection. Infectious virus was demonstrated by using an infectious center assay on Vero cell monolayers. Lymphoid cells isolated from both primary and secondary lymphoid organs also carried infectious virus. The number of infectious centers varied considerably from animal to animal, being as high as 10,000/106 and as low as 1 to 10/106 viable mononuclear cells. However, within any individual virus carrier mouse the numbers of infected peripheral blood mononuclear cells were found to be constant. Virus was not present in a fully infectious form either on the cell surface or within the cell. Mononuclear cells replicating LCMV were found in the bone marrow-derived (B) cell, thymus-derived (T) cell, and phagocytic, adherent cell populations.

Infectious centers were also demonstrated in the peripheral blood and splenic mononuclear cells of adult mice acutely infected with LCMV. The levels of such cells rose to a peak of 100/106 mononuclear cells by day 3 to 4 after i.p. inoculation with virus and then decreased to below detectable levels (<1/106) by day 8.

These results indicate that after infection in utero or at birth, virus may replicate in cells of the immune system. Such infected lymphoid cells may act as a reservoir of infection throughout the animal's life.
Induction and Mechanism of Tolerance to Bovine Serum Albumin in Mice Given Total Lymphoid Irradiation (TLI)
Authors: Zan-Bar Israel , Slavin Shimon , Strober Samuel
Published: December 2022

BALB/c mice given total lymphoid irradiation (TLI) were injected i.p. with bovine serum albumin (BSA) in saline, and challenged with DNP-BSA in complete Freund's adjuvant 6 weeks later. The latter animals made no anti-DNP antibody response as measured by a modified Farr assay, but made a normal anti-DNP response after challenge with DNP-BGG in adjuvant. Normal mice or mice given whole body irradiation were not tolerized by the i.p. injection of BSA in saline. Spleen cells from unresponsive mice (TLI + BSA in saline) suppressed the adoptive secondary anti-DNP response of sublethally irradiated syngeneic hosts given BSA-primed T cells, DNP-BSA-primed B cells, and DNP-BSA in saline. The suppressor cells were antigen specific, and were inactivated by in vitro treatment with anti-Thy 1.2 antiserum and complement. The findings suggest that soluble antigens administered to mice after TLI evoke a state of tolerance that is maintained by antigen-specific suppressor T cells. A similar mechanism may be involved in the maintenance of tolerance to allografts. These findings may have important clinical implications for patients treated with TLI for lymphoid malignancies.
Myeloma-Induced Immunosuppression: A Multistep Mechanism
Authors: Katzmann Jerry A.
Published: December 2022

Hosts of plasma cell tumors have a depressed primary antibody response. We have investigated the ability of cell homogenates and culture fluids from short-term cultures of spleen cells and tumor cells of mice bearing the MOPC-315 plasmacytoma to suppress the in vivo primary antibody response to sheep red blood cells. The homogenates and culture fluids of both MOPC-315 spleen cells and tumor cells suppress the antibody response in a dose-dependent manner. Culture fluids of spleen cells from tumor-bearing mice contain a 10,000- to 20,000-dalton immunosuppressive factor. Culture fluids of non-adherent tumor cells contain a high m.w. suppressor. Injection of the high m.w. tumor suppressive factor into normal mice induces the expression or appearance of host cells that secrete the 10,000- to 20,000-dalton immunosuppressor. The tumor suppressive factor, but not the spleen factor, causes an alteration of lymphocyte membranes such that the anti-DNP activity of the MOPC-315 myeloma protein can be detected on the circulating lymphocytes of injected mice.
In Vitro Growth of Cytotoxic Human Lymphocytes
Authors: Strausser John L. , Rosenberg Steven A.
Published: December 2022

Human lymphocytes sensitized to allogeneic determinants in vitro were continuously cultured on medium prepared from phytohemagglutinin- (PHA-P) stimulated human mixed lymphocyte cultures. With such human-conditioned medium (HCM), human peripheral lymphoid cells could be grown in culture for over 90 days with a doubling time of about 54 hr. Cytotoxic lymphocytes could be grown in culture for this time with no loss of cytotoxicity or change in cytotoxic specificity.
Anti-Idiotypic Antisera in Man
Authors: Geha Raif S. , Weinberg Robert P.
Published: December 2022

Antisera were raised in rabbits against immunosorbent-purified F(ab′)2 fragments of IgG antitetanus toxoid (TT) antibodies obtained from three different donors. The antisera were rendered idiotype specific by absorption with insolubilized TT-nontreactive F(ab′)2. The resulting anti-idiotypic antisera precipitated more than 85% of 125I-radiolabeled F(ab′)2 anti-TT and were shown to be individual specific in that they reacted only with the immunizing F(ab′)2 anti-TT and not with F(ab′)2 anti-TT derived from other donors. Anti-idiotypic antiserum inhibited binding of 125I-TT but not of 125I-DT to IgG derived from the donor of the immunizing F(ab′)2 anti-TT, but did not inhibit 125I-TT binding to IgG derived from other donors. Binding of radiolabeled TT to IgM and IgE was significantly inhibited by the anti-idiotypic antiserum, demonstrating the presence of shared idiotypic determinants on anti-TT antibody molecules belonging to the IgG, IgM, and IgE classes of immunoglobulin. Finally, binding of 125I-F(ab′)2 anti-TT to the antiidiotypic antiserum was completely inhibited by IgG derived from the same donor but only partially inhibited by a great excess of TT antigen, suggesting that the antiidiotypic antiserum is recognizing nonantigen-binding idiotypic determinants on F(ab′)2 anti-TT. The data presented demonstrate that anti-idiotypic heteroantisera can be successfully raised against human antibodies to TT, indicate minimal cross-reactivity of idiotypic determinants between unrelated individuals, and suggest the presence of nonantigen binding-idiotypic determinants on the antibody molecule.
A General Method for the Isolation of the VH Domain from IgM and Other Immunoglobulins
Authors: Rodwell John D. , Karush Fred
Published: December 2022

A chemical cleavage procedure has been developed for IgM, which leads to the isolation of a fragment (Vµ) consisting of the VH region and a small number of N-terminal residues from the Cµ1 domain. The sulfhydryl groups resulting from the reduction of the L-H disulfide linkage are converted to the S-cyano form by treatment with 2-nitro-5-thiocyanobenzoic acid. Cleavage of the H chain on the amino side of the cyanylated residue is effected by incubation overnight at pH 9 and 37°C. The Vµ fragment is the only one-domain fragment formed and is separated from the other products by gel chromatography with Sephacryl S-200 in 6 M guanidine-HCl. The Vµ fragment has been identified by its C-terminal residues on the basis of the established sequence of the human µ-chain. Additional characterization and verification was provided by isoelectric focusing and gel chromatography.

The cleavage procedure is probably generally applicable to IgM of all species. This conclusion is based on the anticipation that the decisive structural feature of the disulfide linkage of the L chain to a half-cystine located near the N-terminal end of the Cµ1 domain is a general feature of IgM structure. Since this arrangement of the L-H bond also is found in other classes and subclasses of immunoglobulin isolation of VH-containing fragments is probably feasible in these instances. Among the many uses of Vµ (and other similar VH-containing fragments) are the preparation of anti-idiotype antibody specific for VH determinants, the evaluation of the contribution of VH to the binding of ligand, and the identification of the VH domain on B and T lymphocytes with antisera specific for the framework determinants of VH.
T Cell Specific Ia Antigens of the I-J and I-E Subregions of H-2
Authors: Hayes Colleen E. , Bach Fritz H.
Published: December 2022

(B10 × B10.D2)F1 mice were immunized with B10.A(5R) concanavalin A-stimulated thymocyte blasts. The genetic disparity between donor and recipient was restricted to the I-J and I-E subregions of the H-2 complex. A high-titered, T cell specific, anti-I-JkEk serum was obtained. The antiserum lysed 27–30% of haplotype k, q, or s lymph node cells, 5.3 ± 2% of haplotype k spleen cells, and did not lyse thymocytes (<3%). Nylon wool-passed lymph node or spleen cells (H-2k) showed considerable reactivity with anti-I-JkEk serum (35–40% lysis); anti-Thy 1.2 plus complement-treated spleen cells did not react (< 5% lysis). I-Ek antibody was detected by B10.A(3R) lymph node cell reactivity (20 ± 3%), whereas reaction with H-2k lymph node cells following B10.A(3R) absorption demonstrated antibody to an I-Jk determinant(s) (12 ± 3%). Reciprocal absorption studies indicated that haplotypes q and s cross-react with k by virtue of anti-I-Jk antibody rather than anti-I-Ek antibody.
Presence of a Non-polymorphic Polypeptide Chain in Ia Immunoprecipitates
Authors: Jones Patricia P. , Murphy Donal B. , McDevitt Hugh O.
Published: December 2022

All immunoprecipitates prepared from NP-40 extracts of 35S-methionine-labeled mouse lymphocytes using anti-Ia alloantisera contain a common polypeptide chain, MW 31,000 daltons. The electrophoretic mobility of this molecule in two-dimensional polyacrylamide gel electrophoresis is the same for Ia antigens coded for by loci in the I-A and I-E subregions and for molecules obtained from mice of different H-2 haplotypes and genetic backgrounds. Immunoprecipitates obtained with anti-H-2K, anti-H-2D, and anti-immunoglobulin antisera do not contain this polypeptide chain. This molecule therefore appears to be a nonpolymorphic protein unique to Ia immunoprecipitates. Because it is nonpolymorphic, this chain probably is not recognized by alloantisera and hence may be precipitated only because it is complexed to polymorphic Ia proteins. Thus the occurrence of this invariant chain with Ia antigens appears to be analogous to that of β2 microglobulin, a nonpolymorphic polypeptide chain coprecipitated with H-2K and H-2D antigens.
Recessive Ir Gene in the Major Histocompatibility Complex (MHC) Regulating High Antibody Responsiveness to TNP-MSA
Authors: Urba Walter J. , Hildemann William H.
Published: December 2022

The antibody response to the 2,4,6-trinitrophenyl hapten (TNP) conjugated to autogenous mouse serum albumin (MSA) is regulated by an Ir gene(s) located within the major histocompatibility complex (MHC). Both the qualitative and quantitative ability of congenic strains to produce TNP-specific antibodies are functions of the H-2 haplotype. Thus, mouse strains may be classified as high (H-2d), intermediate (H-2b&s), and low responders (H-2a,k,n,p&q). Antibody responses measured by antigen-binding capacities in modified Farr assays were compared among many strains carrying recombinant H-2 haplotypes and their hybrid progenies. The gene locus controlling specific responsiveness to TNP-MSA, now designated Ir-6, was mapped within the I-B subregion of the H-2 complex. Recessive inheritance of high responsiveness was confirmed in hybrid progenies of three different low × high responder crosses. IgG1 and IgM TNP-antibody levels of high and low responder mice were separately determined by solid phase radioimmunoassay using class-specific antisera.
Genetic Control of T-Lymphocyte Proliferative Response to Sperm-Whale Myoglobin and Its Synthetic Antigenic Sites in Mice
Authors: Okuda Kenji , Twining Sally S. , Atassi M. Zouhair , David Chella S.
Published: December 2022

Previously we have reported the precise determination of the entire antigenic structure of sperm-whale myoglobin. This protein which is made up on a single polypeptide chain (153 residues, m.w. 17,816) has five antigenic sites located in exposed conformationally sensitive continuous portions of its polypeptide chain. The five antigenic sites occupy the following regions: Site 1: sequence 15–22; Site 2: sequence 56–62; Site 3: sequence 94–99; Site 4: sequence 113–119; Site 5: sequence 145–151. We investigated the genetic control of T lymphocyte proliferative response to myoglobin and its synthetic antigenic sites in mice (PETLES Assay). Two parts of the myoglobin molecule (sequences 1–6 and 121–127) that were not within antigenic sites were also synthesized and employed as controls. Also used as control was a synthetic ‘nonsense’ heptapeptide that did not resemble any part of myoglobin. Haplotypes H-2d, H-2f and H-2s gave substantial T lymphocyte proliferative response to whole myoglobin whereas haplotypes H-2d, H-2k, H-2p, H-2q and H-2r were low responders.
Relationship Between Fc Receptors and Ia, or Mls-Associated Antigens on the Surface of B Lymphoma Lines
Authors: Kanellopoulos-Langevin Colette , Kim Kyung Jin , Sachs David H. , Asofsky Richard
Published: December 2022

A panel of spontaneous BALB/c B cell lymphoma lines with different cell surface phenotypes were used to study the relationship of Fc receptors, Ia and Mls-associated (tentatively called LyM) antigens. These cultured lymphomas have the following B cell characteristics: They (a) express surface Ig; (b) have Fc receptors tested by sheep erythrocyte-antibody rosetting assay; (c) do not ingest latex particles, and (d) are negative for esterase activity.

One of these cell lines, A-20, is Ia+, Fc+, LyM+; another, M 12, is Ia-, Fc+, LyM+. Using these and other lines, we have shown that Fc receptors on Ia+ cells (e.g. A-20) can be inhibited by anti-Ia sera. There is no such inhibition on Ia- cells (e.g. M 12). However, anti-Mls serum can inhibit Fc rosetting on tumors with either Ia+ or Ia- phenotype.

Thus we can demonstrate inhibition of Fc receptors with anti-Ia sera similar to that reported for normal spleen cells.
The Effect of Anti-Ia Alloantisera on Syngeneic Tumor Immunity
Authors: Greene Mark I. , Perry Linda , Dorf M. E. , Benacerraf Baruj
Published: December 2022

Tumor specific suppressor T cells (STC) arise in A/J mice as a consequence of inoculation of syngeneic methylcholanthrene induced fibrosarcomas, S1509a and SAI, and elaborate tumor antigen specific suppressor factor(s) (SF). SF which bears determinants encoded by the K end of the H-2 MHC, and STC which express I-J subregion encoded cell surface determinants dampen the primary response and limit the secondary effector reactivity to tumor in vivo. Daily treatment of normal A/J mice with 2 µl of (DBA/2 × 3R)F1 anti-5R (anti-I-J) antisera beginning at the time of a 106 S1509a tumor inocula resulted in a highly significant reduced tumor growth and an abrogation of the capacity of such mice to adoptively transfer suppression. Furthermore, anti-I-J antisera could also limit tumor incidence and appearance when used with smaller tumor inocula. The sera effect was specific and was lost only after absorption with lymphoid cells of the appropriate haplotype.
Antibody Responses to GAT by (Responder × Nonresponder)F1 Spleen Cells Stimulated by Parental GAT-Macrophages
Authors: Pierce Carl W.
Published: December 2022

The development of IgG PFC responses specific for the random terpolymer of l-glutamic acid60-l-alanine30-L-tyrosine10 (GAT) by virgin and immune (responder C57BL/10 × nonresponder DBA/1)F1 spleen cells after stimulation with responder and nonresponder parental or F1 GAT-pulsed macrophages (GAT/Mϕ) in vitro was investigated. Virgin F1 spleen cells developed comparable primary responses to GAT-MBSA and to both parental and F1 GAT-Mϕ. F1 spleen cells from mice immunized with responder or nonresponder parental GAT-Mϕ developed secondary responses only after stimulation with the parental GAT-Mϕ used for immunization or F1 GAT-Mϕ. These observations are consistent with previous observations that spleen cells from mice immunized with syngeneic or allogeneic GAT-Mϕ develop secondary responses only with GAT-Mϕ syngeneic at the I-A subregion of the H-2 complex with the M-ϕ used for priming. By contrast, F1 spleen cells from mice immunized with GAT-MBSA or F1 GAT-Mϕ developed secondary responses after stimulation with either parental or F1 GAT-Mϕ.
Primary in vivo Immunization of F1 (2 × 13) Guinea Pigs by Parental Macrophages Bearing Multideterminant and Genetically Restricted Antigens
Authors: Yokomuro Kozo , Rosenthal Alan S.
Published: December 2022

An objection has been raised to the concept of macrophage Ir gene function by the in vivo experiments of Pierce et al. in which nonresponder murine macrophages (Mϕ) primed responder mice, thus suggesting Ir gene function at the level of the T cell. Both to examine this issue in the guinea pig and to determine whether in vitro secondary responses are solely reflections of the animal's initial immunologic experience, inbred strain 2 and 13 guinea pigs were immunized in vivo with syngeneic or allogeneic antigen bearing Mϕ. Monolayer purified Mϕ were pulsed with oxidized B chain of insulin and dinitrophenylated ovalbumin (DNP-OVA) for 60 minutes, washed and 2 × 107 injected intradermally into recipient animals. Fourteen days later, in vitro antigen induced specific T cell proliferation was assessed. When strain 13 guinea pigs, high responder to both antigens, were immunized by antigen pulsed strain 13 Mϕ, T cells proliferated in vitro to both antigens.
H-2 Restriction of the Activity of Allogeneic Effect Factor
Authors: Delovitch Terry L. , Sohn U.
Published: December 2022

The helper activity of allogeneic effect factors (AEF) derived from mixed lymphocyte culture reactions (MLR) between activated responder cells and either I-region, I-subregion, or Mls-region incompatible irradiated stimulator spleen cells was analyzed. An AEF generated across either an a) entire I-region, b) I-A, I-B and I-J, c) I-J or d) I-E and I-C subregion incompatibility helps B cells of only the stimulator haplotype, or of haplotypes which carry I-subregions selectively expressed by the stimulator cells and not the responder cells. An AEF generated across an Mls-region difference provides weak help for B cells only of haplotypes which share both an Mls and I-region identity with the stimulator haplotype. The observed lack of help for B cells of the responder haplotypes is not due to a suppressor factor activity in these H-2 restricted AEFs. These results are consistent with those obtained for another genetically restricted AEF produced by Ia negative activated responder cells and H-2 incompatible irradiated T-cell-depleted stimulator cells.
Mouse Natural Killer Cells: Induction Specificity, and Function
Authors: Welsh Raymond M.
Published: December 2022

Attention has recently focused on the possible significance of natural killer (NK)2 lymphocytes in the surveillance of tumors by the immune system (1). This is because NK cells from several species preferentially lyse transformed target cells in vitro (2–5). NK lymphocytes from mouse and man have both been studied in detail, but the manipulability of the mouse model has allowed for the better resolution of NK cell properties. In this review I will restrict most of my comments to the mouse NK cell system. The properties of human NK cells have recently been reviewed elsewhere (6).

Characterization and tissue distribution. It is agreed that mouse NK cells are x-ray-resistant (7) lymphocytes lacking cell surface immunoglobulin (8, 9). They are thought to be derived from bone marrow since their activity is removed by depletion of bone marrow cells with anti-bone marrow antibody or with 89Strontium (7, 10).
Relation of the Urinary Cancer-Related Glycoprotein EDC1 to Plasma Inter-α -Trypsin Inhibitor
Authors: Chawla Rajender K. , Wadsworth Allan D. , Rudman Daniel
Published: December 2022

Previous studies showed 30 to 60% of patients with disseminated neoplastic disease excrete in their urine 100 to 500 mg/day of a novel glycoprotein, m.w. 27,000, labeled EDC1. Normal urine contains <1 mg/day. Both normal and cancer plasmas contain a large m.w. factor that cross-reacted with antiserum to EDC1.

Immunoelectrophoresis (IEP) of normal plasma versus rabbit antiserum to EDC1 shows one precipitin line that is of slower mobility than EDC1. Gel filtration of plasma on Sephadex G-150, and analysis of the effluent by double immunodiffusion (DID), IEP, and radioimmunoassay (RIA) with antiserum to EDC1, reveal that the plasma EDC1-immunoreactive component has a m.w. of about 150,000.

Among antisera to 45 purified plasma proteins, only that to inter-α trypsin inhibitor (IATI) m.w. 170,000, reacts with EDC1. This finding suggests that the large EDC1-immunoreactive component is IATI. This is confirmed by the following observations with purified IATI: EDC1 and IATI give a reaction of partial identity in DID versus anti-EDC1 and versus anti-IATI; IATI and plasma give a line of identity in DID versus anti-EDC1; IATI and the immunoreactive component in plasma have the same mobility in IEP versus anti-EDC1. Furthermore, EDC1 possesses an antiproteolytic property closely similar to that of IATI (both proteins inhibit trypsin and chymotrypsin but not plasmin, pepsin, elastase, or exopeptidases).

We conclude that cancer-related urinary glycoprotein EDC1 is related antigenically and chemically to IATI and may be a cleavage product of the latter.
Characterization of Immune Complexes in Serum by Adsorption on Staphylococcal Protein A: Model Studies and Application to Sera of Rats Bearing a Gross Virus-Induced Lymphoma
Authors: Tucker Derek F. , Begent Richard H. J. , Hogg Nancy M.
Published: December 2022

A procedure to isolate soluble IgG-containing immune complexes from serum was developed, which combined Sephadex G-150 filtration with affinity chromatography on Staphylococcal protein A-Sepharose. The effectiveness of the method was shown by isolating model immune complexes of 125I bovine plasma albumin (BPA) and antibody to BPA from serum. Functionally intact antibody and antigen were recovered from these complexes by chromatography on Sephadex G-150 at low pH and the degree of purification was demonstrated by NaDod.SO4 polyacrylamide slab gel electrophoresis (PAGE) and autoradiography of the immune complexes and their components.

Application of these methods to a Gross virus-induced rat lymphoma model tested their potential for characterizing naturally occurring circulating immune complexes (CIC). Sera from these animals were shown by Raji cell immunofluorescence to contain high concentrations of CIC 15 to 21 days after tumor inoculation. CIC could be isolated from these sera by the procedures developed for model complexes. Analysis by PAGE indicated that they contained primarily a protein of apparent m.w. 79,000 complexed with IgG. This putative antigen component of CIC (79,000 daltons) reacted with a rabbit antiserum directed against the murine leukemia virus (MuLV) envelope glycoprotein, gp70. Techniques using protein A affinity chromatography may also be applicable to characterization of both antigen and antibody components of CIC in human diseases including cancer.
A Subpopulation of Adherent Accessory Cells Bearing Both I-A and I-E or C Subregion Antigens Is Required for Antigen-Specific Murine T Lymphocyte Proliferation
Authors: Cowing Carol , Pincus Seth H. , Sachs David H. , Dickler Howard B.
Published: December 2022

A murine T lymphocyte proliferation assay that used antigen-primed lymph node T cells, was antigen specific, and required exogenous accessory cells was used to characterize the accessory cells that supported proliferation. These cells were Thy 1.2 negative, radioresistant, glass-adherent, and were functional only if alive. The accessory cell function of spleen adherent cells was much greater than that of peritoneal cells. Also, the accessory cell function of spleen adherent cells was proportional to the length of time such cells were incubated with antigen and very small numbers of such cells provided accessory cell function. Cytotoxic studies with subregion-restricted anti-Ia antibodies and complement indicated that accessory cell function resided in a subpopulation of spleen adherent cells that bore both I-A and I-E or C subregion antigens. The function of such cells was not related to a selective ability (vs other spleen adherent cells) to take up antigen. These data indicate that antigen-specific stimulation of T lymphocyte proliferation requires at least one specific subpopulation of spleen adherent cells that can be phenotypically identified by its expression of Ia antigens and are consistent with the possibility that Ia antigens may be Ir gene products.
Role of Prostaglandin Synthesis in Rabbit Platelet Activation Induced by Basophil-Derived Platelet-Activating Factor
Authors: Shaw J. O. , Printz M. P. , Hirabayashi K. , Henson P. M.
Published: December 2022

Rabbit platelet-activating factor (PAF) is a lipid released from IgE-sensitized basophils and mast cells during challenge with antigen. PAF induces shape change, aggregation, and secretion of granule-associated 3[H]-serotonin in rabbit platelets. This study investigated the capacity of PAF to initiate rabbit platelet prostaglandin synthesis and the requirement for generated prostaglandin endoperoxides and thromboxane A2 in PAF-induced platelet aggregation and secretion. PAF in submaximal concentrations induced synthesis in rabbit platelets of PGE2, PGF2α, and prostaglandin endoperoxides in amounts comparable to that initiated by collagen, but greater than that caused by thrombin. PAF stimulation of platelets preloaded with 14[C]-arachidonic acid caused platelet production of 14[C]-thromboxane B2, the stable derivative of 14[C]-thromboxane A2. When compared kinetically to secretion, PAF induced platelet endoperoxide production peaked just before maximal release of granular 3[H]-serotonin content. However, the cyclooxygenase inhibitors, aspirin and indomethacin, failed to alter PAF-induced platelet shape change or aggregation. Similarly, secretion was unaffected by indomethacin except for a slight attenuation seen at the lowest stimulus concentrations examined. Platelet production of endoperoxides occurred throughout the range of PAF concentrations examined, even those causing barely detectable levels of secretion. PAF thus initiates platelet synthesis of prostaglandin endoperoxides and thromboxane A2, but aggregation and secretion induced by PAF occur independent of these products of the cyclooxygenase pathway.
Requirement for an Ia-Bearing Accessory Cell in Con A-Induced T Cell Proliferation
Authors: Ahmann Gerald B. , Sachs David H. , Hodes Richard J.
Published: December 2022

Preatment of murine lymphoid cells with anti-Ia and C abrogated the proliferative response of these cells to Con A, but not to PHA. Reconstitution experiments demonstrated that T cell-enriched populations failed to restore Con A responsiveness and that T cell-depleted populations were more effective in restoring responsiveness to Con A. In particular, a population of 1000 R resistant, glass-adherent, non-T spleen cells was capable of completely restoring responsiveness to Con A when added in numbers as low as 4% of cultured cells. These splenic adherent cells were found to express Ia determinants encoded by at least two genes: one in I-A and the other in I-B, I-J, and/or I-E/C, and it was demonstrated that determinants encoded in these two regions were expressed on the same cell. These results demonstrate that non-T accessory cells may be the Ia+ cells entirely responsible for the anti-Ia and C-induced abrogation of T cell proliferative responses to Con A.
Studies on Immunosuppression by Cobra Venom Factor
Authors: Matsuda T. , Martinelli G. P. , Osler A. G.
Published: December 2022

We have reassessed the effects of CVF administration on the humoral responses to two T-independent immunogens: DNP·Ficoll and DNP·Polyacrylamide. With high immunogen doses, little or no evidence of suppression was found. However when the immunizing dose was reduced, suppression of both IgG and IgM responses became apparent. As indicated in a previous report, the immunosuppressive effect of CVF on T-dependent responses may result not only from C depletion but also from the generation of C cleavage products that may impair the auxiliary contribution of macrophages to the generation of these humoral responses. A similar mechanism may be applicable to the suppression of antibody production to DNP·Ficoll and DNP·PAA in view of recent reports showing a macrophage requirement for the response to these immunogens.