bluuebunny/crossref_metadata_2025 huggingface dataset:
Lymphocyte Function in Experimental African Trypanosomiasis
Authors: Mansfield John M. , Bagasra Omar
Published: December 2022

In order to obtain a better understanding of immune system dysfunction in chronic African trypanosomiasis, we have assessed primary B cell responses to helper T cell-independent and -dependent antigens in C57BL/6 mice infected with the human pathogen, Trypanosoma rhodesiense. Mean survival time of infected mice was 38 days. All animals exhibited fluctuating parasitemias until 5 to 7 days before death when parasitemias remained high. Thymus size decreased as infection progressed, but spleen size increased up to 20 X normal. The proportion of macrophages (nonspecific esterase-positive cells) and B cells or B cell progeny (Ig+) increased in spleen, whereas the proportion of T cells (θ+) decreased. However, in a terminal mouse with enlarged spleen, the absolute increase in total cell numbers reflected a 1.7 X increase in T cells, an 11.2 X increase in B cells, and a 21 X increase in macrophages compared to controls. The ability of infected mice to mount primary in vivo splenic PFC responses to helper T cell-independent antigens (SIII, PVP) was elevated throughout most of infection when compared to control animals responses; serum antibody responses to SIII or PVP paralleled the enhanced PFC responses. The terminal phase of infection was marked by a failure of B cells to respond to these antigens. The elevated primary splenic PFC responses observed during infection to T-independent antigens such as SIII may be attributed to loss of suppressor T cell control, since infected mouse spleen cells did not respond to low-dose priming with SIII. Primary in vivo B cell responses to helper T cell-dependent antigens (SRC, TNP-KLH) were initially elevated when antigen and parasites were given on the same day, but detectable primary splenic PFC responses to these antigens could not be induced after several days of infection. Suppression of PFC responses to TNP-KLH in infected mice could be completely abrogated if helper T cells were carrier-primed in animals before infection. On the basis of our studies, we suggest that intrinsic B cell function is intact and elevated throughout most of infection, but that a terminal failure of B cells precedes death. We further suggest that a loss of function of accessory and regulatory cells (helper and suppressor T cells) may be responsible for the irregularities in B cell function observed during chronic African trypanosomiasis.
Inhibition of Specific Effector T Cell-Target Cell Conjugates by Target Cell Plasma Membranes
Authors: Gilmer Penny J. , McDevitt Hugh O. , McConnell Harden M.
Published: December 2022

Conjugate formation between effector T cells raised in vivo in a BALB/c host and fluorescein diacetate-labeled allogeneic EL4 tumor target cells was observed and scored by using a fluorescence microscope. This conjugate formation was found to be specifically inhibited by plasma membrane vesicles isolated from the EL4 cells.
Suppression of Growth of Guinea Pig Line-10 Hepatocarcinoma
Authors: Brunda Michael J. , Minden Percy , Heusser Christoph H. , Ferguson H. Rodney
Published: December 2022

Suppression of growth of the line-10 hepatocarcinoma in strain-2 guinea pigs occurred when line-10 cells were injected intradermally together with sera or immunoglobulins derived from normal rabbits. A significant number of animals were resistant to subsequent rechallenge with tumor cells. This immunity was specific, depended on contact of immunoglobulins with tumor cells and on the concentration of immunoglobulins. Repeated injections acted as potent vaccines and resulted in the development of immunity in 84.6% of recipients. Fc receptors were not detected on line-10 cells. Antibodies reacting with line-10 cell unique antigens as well as with antigens common to line-10, line-1 and normal guinea pig spleen cells were found in NRS. Injection of line-10 cells together with rabbit immunoglobulins from which antibodies reacting with antigens derived from line-10 cells had been removed did not resulted in tumor suppression. The specific antigen(s) recognized by antibodies that suppressed growth of the line-10 tumor in vivo was not determined.
Reaginic Antibody Formation in the Mouse
Authors: Tamura Shin-Ichi , Ishizaka Kimishige
Published: December 2022

The kinetics of IgE antibody response to alum-absorbed dinitrophenyl derivatives of ovalbumin (DNP-OA) was dependent on the dose of immunogen. A persistent IgE antibody response was obtained when high responder BDF1 mice were immunized with a minimum (0.05 µg) dose. An increase of the immunogen to 10 µg depressed IgE antibody responses but enhanced IgG antibody responses of both hapten and carrier specificities. Determination of T helper cell activity and B memory cells after immunization with different doses of antigen indicated that minimum immunogen was favorable for developing helper activity, whereas 1 to 10 µg immunogen were more favorable than a 0.05-µg dose for developing both IgE and IgG B memory cells. Nevertheless, neither helper T cells nor B memory cells in the spleen explains a transient IgE antibody response to a high (10 µg) dose of DNP-OA. Evidence was obtained that immunization with 10 µg OA induced generation of antigen-specific suppressor T cells, which were not detectable after immunization with 0.05 µg OA. Transfer of suppressor T cells to DNP-OA-primed mice depressed both anti-hapten and anti-carrier IgE antibody responses. The results suggested strongly that suppressor T cells are involved in a transient IgE antibody response to a high-dose immunogen.
Tolerance Induced by TNP-Derivatized Syngeneic Erythrocytes: Evidence for Cooperation between Hapten-Specific T and Hapten-Specific B Lymphocytes in the Immune Rensponse
Authors: Moody Charles E. , Innes Judith B. , Siskind Gregory W. , Weksler Marc E.
Published: December 2022

Tolerance to the DNP haptenic determinant was induced with a single i.v. injection of trinitrophenylated syngeneic red blood cells. The tolerant state lasted 1 month and was stable on transfer to irradiated thymectomized syngeneic recipients. Suppressor activity was found soon after injection of tolerogen but was lost before the termination of tolerance. The unresponsive state could be reversed by adding normal thymus cells to tolerant spleen cells but not by normal bone marrow cells. LPS when given with immunogen restored the normal immune response in tolerant mice.

Thus the injection of TNP-MRBC induced partial immune unresponsiveness which was characterized by the induction of T cell suppressor activity and by a hapten-specific helper T cell tolerance. Finally, these studies suggest a cooperative interaction between DNP-specific T lymphocytes and DNP-specific B lymphocytes in the immune response to DNP-BGG.
Intrinsic Tritium Labeling of the Capsular Polysaccharide Antigen of Haemophilus Influenzae Type B
Authors: Anderson Porter
Published: December 2022

The capsular polysaccharide of Haemophilus influenzae type b was intrinsically labeled with tritium by a microculture technique with 6-3H-d-glucose and was isolated in radioantigenically pure form by a combination of selective precipitation and molecular sieve chromatography. Labeling with tritated sugar residues approached one-fourth maximum and produced a specific activity 10-fold that previously described for extrinsic labeling methods. In radioantigen-binding assays for antibody, sensitivity depended on the size of the antigen; preparations were readily made that could detect 0.01 µg Ab/ml in serum samples of 25 µl. Stability of the labeled antigen appears limited only by the primary radiodecomposition of tritium.
Antigen-Initiated B Lymphocyte Differentiation
Authors: Howard Maureen C. , Fidler John M. , Hamilton John , Shortman Ken
Published: December 2022

The effects of injection of unrelated antigens on the sedimentation velocity and buoyant density characteristics of specific “virgin” or “memory” adoptive transfer AFC-progenitors in unprimed or long-term hapten-primed adult mouse spleen were examined to determine if the physical properties of these B cells could be altered by nonspecific stimulation. Spleen cells from normal unprimed adult CBA mice (as a source of virgin B cells) or from long-term hapten-primed CBA (as a source of memory B cells) were separated and the fractions were tested for NIP-specific AFC-progenitor activity with an adoptive immune assay system that gave a direct linear measure of B cell activity. If the donor mice were injected 1 day previously with POL or HRC, the NIP-specific virgin IgM AFC-progenitor activity shifted from medium sized, medium-light density lymphocytes to larger, much lighter cells. The same 1-day nonspecific stimulation had little effect on the physical properties of either IgM or IgG memory AFC-progenitors, although a small increase in the size of such cells was observed 2 days post immunization with unrelated antigens. Nonspecific stimulation caused identical physical changes in the virgin AFC-progenitors of athymic nude mice, but only partially altered the physical properties of virgin AFC-progenitors in neonatal mice. In the latter case, a normal transformation occurred if adult, radioresistant macrophages were injected with the stimulating antigens.

These results indicate that a particular subset of virgin B cells responds nonspecifically to antigenic injections, becoming larger cells of lighter density. The mechanism of this effect appears to involve mature macrophages, but not functional T lymphocytes. Memory B cells do not readily respond in this way.
Electrophoretic Mobility Distributions of Normal Human T and B Lymphocytes and of Peripheral Blood Lymphoblasts in Acute Lymphocytic Leukemia: Effects of Neuraminidase and of Solvent Ionic Strength
Authors: Smith Barton A. , Ware B. R. , Yankee Ronald A.
Published: December 2022

Human T and B lymphocytes were found to be distinguishable on the basis of electrophoretic mobility, with the T cells having the higher mobility, in agreement with previous reports. The effects of the enzyme neuraminidase on the electrophoretic mobilities of T and B lymphocytes were determined. T lymphocytes showed a greater decrease in electrophoretic mobility after neuraminidase treatment; the relative mobilities of T and B cells were reversed by neuraminidase treatment, and the electrophoretic distinguishability was enhanced. The electrophoretic mobility distributions of lymphoblasts from patients with acute lymphocytic leukemia were found to differ from those of normal cells in their response to neuraminidase treatment and to changes in solution ionic strength. These results imply that the surface structure of the leukemic cells differs from that of either T or B lymphocytes from normal donors.
Aging and the Immune Response
Authors: Gardner Ian D. , Remington Jack S.
Published: December 2022

Studies were performed to determine the effect of aging on the antibody response and cyst formations after infection with a relatively avirulent strain of the intracellular parasite Toxoplasma gondii. When compared with young mice (4 months), aged mice showed a significant decrease in the magnitude of humoral immune response to infection. This decrease was observed at the peak of the acute infection and also during chronic infection. Evaluation of the presence of Toxoplasma cysts, a measure of the latent infection, revealed that the numbers of tissue cysts present 11 weeks after infection increased with the age of the mice at time of infection. The larger numbers of cysts in older mice which had received the same inoculum size of T. gondii as young mice, together with our previous observations of increased susceptibility of these older mice to T. gondii, suggest that an age-related decrease in the early immune response to this infection allows an increased multiplication of the organism in vivo, leading to increased cyst numbers or death.
Opsonic Activity of Guinea Pig Serum Is Reduced by in Vivo Administration of Carcinogenic Nitrosamines
Authors: Marom Zvi , Brade Volker , Colten Harvey R.
Published: December 2022

The administration of each of four carcinogenic nitrosamines to normal guinea pigs resulted in decreased opsonic activity of their sera but did not affect the capacity of their peritoneal macrophages to phagocytize particles opsonized with normal serum. Diphenylnitrosamine, a noncarcinogenic analogue, had no significant effect on opsonic or phagocytic activity.
Rodent Malaria: BCG-Induced Protection and Immunosuppression
Authors: Smrkovski Lloyd L. , Strickland George T.
Published: December 2022

One dose of 107 viable units of Mycobacterium bovis, strain BCG, protected a significant number of Swiss mice from a primary challenge with 104 thoracic sporozoites of Plasmodium berghei. Immunization with irradiated sporozoites induced greater protection than that observed in BCG-treated animals. Mice treated with BCG and surviving a primary sporozoite challenge were not protected from rechallenge, whereas mice immunized with irradiated sporozoites and surviving initial challenge of sporozoites were solidly immune to further challenge. Immunizing mice with BCG and irradiated sporozoites simultaneously resulted in a synergistic effect of increased protection against a primary challenge of sporozoites only if the two immunogens were administered on the same day and if the mice were challenged 1 to 3 days later. Mice given BCG and irradiated sporozoites and surviving a primary challenge of sporozoites were unable to survive rechallenge. BCG given to mice previously immunized with irradiated sporozoites suppressed their protective immunity against sporozoite challenge.
The Effect of Complement Depletion on the Course of Sindbis Virus Infection in Mice
Authors: Hirsch Robert L. , Griffin Diane E. , Winkelstein Jerry A.
Published: December 2022

The course of Sindbis virus infection in 12-day-old BALB/c mice was altered significantly in animals depleted of the third component of complement (C3) by treatment with purified cobra venom factor (CoVF). Although the same percentage of C3-depleted and normal animals died (30%) after the subcutaneous inoculation of 1000 PFU Sindbis virus, the mean day of death was later in C3-depleted mice (8.4 days) than in controls (6.5 days). In addition, morbidity was prolonged in C3-depleted mice. Growth of virus at the inoculation site in the foot was not different; however, viremia was prolonged and the amount of virus in the brain was 1000-fold greater 6 days after infection in C3-depleted animals. These studies demonstrated that complement plays an important role in the host's response to Sindbis virus infection by participating in both beneficial and immunopathologic responses to the infection.
The Fc Receptors of Primary and Cultured Phagocytic Cells Studied with Homogeneous Antibodies
Authors: Diamond Betty , Bloom Barry R. , Scharff Matthew D.
Published: December 2022

Mouse spleen cell-myeloma hybrids were produced that secreted monoclonal antibodies to SRBC of the IgG2a and IgG2b subclass. These homogeneous antibodies were used to examine the Fc receptors for mouse IgG2a and IgG2b on resident and thioglycollate-induced macrophages and on two continuous macrophage-like cell lines (J774.2 and FC-1). By cross-inhibition experiments, we have shown that all four of these cell types have separate receptors for aggregated IgG2a and IgG2b. Both receptors bind heterogeneous rabbit antibody complexed to antigen. Large amounts of soluble proteins do not inhibit the binding of antigen-antibody complexes to either receptor. The rosetting of IgG2a-coated SRBC, but not of IgG2b-coated SRBC, is inhibited by treating all four cell types with trypsin or cytochalasin and is decreased at 4°C. Both the IgG2a and IgG2b receptors will mediate phagocytosis in all four cell types. Mutants of the FC-1 cell line were obtained that can bind both IgG2a and IgG2b antibody-antigen complexes but phagocytize only through the IgG2b receptor. We conclude from these studies that naturally occurring complexes between antigen and mouse IgG2a and IgG2b antibodies are bound to mouse macrophages and phagocytized through functionally distinguishable Fc receptors. The use of homogeneous monoclonal antibodies facilitates such studies and avoids some of the ambiguities that arise when artificially aggregated immunoglobulins or heterogeneous antibodies are used.
Biochemical Relationship of Thymocyte Mitogenic Factor and Factors Enhancing Humoral and Cell-Mediated Immune Responses
Authors: Farrar John J. , Simon Philip L. , Koopman William J. , Fuller-Bonar Janet
Published: December 2022

Supernatants from concanavalin A-induced mouse spleen cells contain soluble factors which: 1) augment the anti-sheep erythrocyte hemolytic plaque-forming cell response of nude mouse spleen cells, 2) facilitate the generation of antigen-specific cytotoxic cells by alloantigen-stimulated thymocytes, and 3) enhance the mitogenic response of thymocytes in the presence or absence of phytohemagglutinin. The uninduced control spleen cell supernatants contained variable but significantly less biologic activity within the different assay systems. The principal enhancing activity (apparent m.w. 35,000 to 38,000) in each of the assay systems was found to be inseparable after a multiple step purification scheme involving ammonium sulfate precipitation, two-cycle gel filtration, hydroxylapatite chromatography, and hydrophobic chromatography on Phenyl Sepharose. These results indicate that either a single factor (designated thymocyte mitogenic factor) augments both humoral and cell-mediated immune responses or, alternatively, the different biologic activities are mediated by different factors that share a common core structure.
Thy-1 Antigenicity Is Associated with Glycolipids of Brain and Thymocytes
Authors: Wang T. J. , Freimuth W. W. , Miller H. C. , Esselman W. J.
Published: December 2022

Glycolipids with Thy-1 activity have been isolated from mouse brain and thymocytes. An immune response assay has been found to be effective in the identification of these glycolipids. The addition of brain or thymocyte glycolipids from AKR (Thy-1.1) or C3H (Thy-1.2) mice to spleen cells resulted in the generation of antibody-forming cells specific for Thy-1.1 or Thy-1.2, respectively. This response was investigated with a modified plaqueforming cell (PFC) assay by using thymocyte target cells in agarose. Incubation of C3H brain glycolipid or Thy-1.2 brain glycoprotein with AKR spleen cells resulted in an anti-Thy-1.2 PFC response when assayed for PFC in a lawn of C3H thymocytes. Controls in which equivalent amounts of AKR glycolipid were added to these cultures resulted in no PFC response. On the other hand when C3H or AKR glycolipid or Thy-1.2 glycoprotein was incubated with C3H spleen cells, the AKR glycolipid induced an anti-Thy-1.1 PFC response in a lawn of AKR thymocytes, but C3H glycolipid and Thy-1.2 glycoprotein elicited no PFC response. These experiments demonstrated the Thy-1 specificity of the glycolipids. Absorption of the antigens with either anti-Thy-1.1 or anti-Thy-1.2 sera before addition to the cultures also demonstrated Thy-1 antigen specificity.

Development of the immune response assay has permitted further purification of Thy-1 active glycolipids and has led to the following conclusions. Brain GMl ganglioside purified by column and thin layer chromatography from AKR or C3H mice contained Thy-1.1 or Thy-1.2 active glycolipids, respectively. The Thy-1.1 or Thy-1.2 active glycolipids could be separated from GMl by a third thin layer chromatography system. The Thy-1.1 and Thy-1.2 glycolipids were estimated to be only a small percentage of the total AKR or C3H brain GMl preparations. Thy-1 active glycolipids isolated from thymocytes had similar thin layer mobility and exhibited Thy-1 specificity in the system described. We suggest that Thy-1 antigenicity lies in carbohydrate structures that are conjugated to either lipid or protein carriers because we observed Thy-1 specificity associated with both glycolipids and glycoprotein.
Macrophage Migration Inhibition Factor: Interactions with Calcium, Magnesium, and Cyclic AMP
Authors: Block Lutz H. , Aloni Beny , Biemesderfer Daniel , Kashgarian Michael , Bitensky Mark W.
Published: December 2022

Human migration inhibition factor (MIF), at concentrations that inhibit guinea pig macrophage migration, do not alter the cytoplasmic levels of cAMP. However, modest elevations of intracellular cAMP, achieved either with isoproterenol or by the addition of db cAMP, markedly interfere with the action of MIF. This effect is, however, very time dependent. Elevation of macrophage cAMP 1 hr after the addition of MIF has no effect on MIF action. The addition of isoproterenol or db cAMP, caused a brief rise of macrophage cAMP levels, and this was followed by a period of unresponsiveness to MIF that lasts for at least 1 hr. In addition, we find that nonphysiologic increases in cAMP (caused by the addition of 1 mM db cAMP) can also interfere with macrophage migration.

We have also studied the dependence of MIF action and binding on divalent cations. The calcium ionophore A23187 (which causes a 3-fold increase in the uptake of calcium by the macrophages) resembles MIF in preventing macrophage migration. However, MIF itself does not increase calcium uptake. Calcium and magnesium do appear necessary, however, for binding MIF to the macrophage cell surface and possibly for the continuing action of MIF.

Finally, examination of the macrophage surface with the scanning electron microscope reveals that the inhibition of migration by MIF, cAMP, or A23187 are all accompanied by different morphologic arrangements of the macrophage cell membrane. We conclude that the inhibition of macrophage migration produced by these three agents depends on different underlying mechanisms.
Guinea Pig Macrophage Agglutination Factor Is Antigenically Distinct from Migration Inhibition Factor and Immunoglobulin
Authors: Godfrey H. P. , Geczy Carolyn L.
Published: December 2022

Lymph node cells from guinea pigs with specific delayed hypersensitivity release macrophage agglutination (MAggF) and migration inhibition factors (MIF) upon exposure to antigen or concanavalin A in serumfree medium. MAggF in culture supernatants was absorbed neither by immunoabsorbents made with a rabbit anti-guinea pig lymphokine serum that removed MIF, nor by immunoabsorbents made with rabbit anti-guinea pig Ig. These results suggest that MAggF is antigenically distinct from MIF and Ig.
Lymphocyte-Activating Factor
Authors: Economou James S. , Shin Hyun S.
Published: December 2022

Lipopolysaccharide-activated murine peritoneal macrophages elaborate lymphocyte-activating factor (LAF) which is mitogenic for murine thymocytes. A method of LAF production is presented that permits the generation of a relatively homogeneous molecular species. LAF has an isoelectric point of 4.8 (range 4.7–4.9). The m.w. was determined by using several physical techniques. The apparent sedimentation coefficient (S20,w) was determined to be 2.0S by sucrose gradient ultracentrifugation. The Stokes (molecular) radius was determined by gel filtration on Sephadex G-75 to be 22 Å (range 21.5 to 22.5); the calculated diffusion coefficient (D20,w) was 9.7 × 10-7 cm2/sec (range 9.5 × 10-7 to 9.9 × 10-7). The buoyant density of LAF is 1.30 g/cm3 (range 1.27 to 1.33) as determined by CsCl isopycnic ultracentrifugation; the partial specific volume was estimated to be 0.72 (range 0.70 to 0.74). From these data, the m.w. was calculated to be 18,000 daltons (range 16,400 to 19,600) with the Svedberg equation. The frictional ratio was calculated to be 1.25.
Evidence for Close Linkage of a Mouse Light Chain Marker with the LY-2,3 Locus
Authors: Gibson David M. , Taylor Benjamin A. , Cherry Marianna
Published: December 2022

Light chains associated with normal serum immunoglobulin can be resolved into a finite number of discrete focusing bands by isoelectric focusing. Four distinct light chain patterns can be distinguished among the inbred mouse strains. In the present studies inheritance of the characteristic light chain patterns has been studied in the AKXL recombinant inbred lines (derived from C57L/J and AKR/J parental lines) and in the inbred Ly-2a,3a congenic line B6.PL-Ly-2aLy-3a/Cy as well as in individual backcross animals of an incipient Ly-2a,3a congenic strain. Virtually complete concordance was observed for the expression of light chains characteristic of phenotype B (AKR/J-like) and the expression of the Ly-2a,3a allele. This observation indicates that a locus controlling light chain structure and/or expression is closely linked (less than 2.6 map units) to the Ly-2,3 locus on mouse Chromosome 6. The locus controlling normal light chain IF-patterns has been designated Ef1.
Regulation of the IgE Antibody Response in Mice
Authors: Peeters Suzanne H. , Carter Brian G.
Published: December 2022

An adoptive cell transfer system was defined for studying the long-term IgE anti-ovalbumin (OA) response of B6D2F1 mice. The primary IgE anti-OA response given by this strain persisted for more than 8 months and the half-life of the IgE anti-OA in circulation was 10.5 hr as measured by residual passive cutaneous anaphylaxis (PCA) in rat skin. The system described for studying cell recruitment patterns in this response involves the transfer of primed spleen cells to irradiated recipients and the production in these recipients of IgE and hemagglutinating antibody without further overt antigenic challenge. The capacity of primed spleen cells to transfer the response was detectable 10 days after immunization and increased for the next 6 weeks. After this time the ability of primed cells to transfer the response declined steadily, but was still evident 8 months after immunization of the donor animals. The transferred response itself persisted in the recipients for many months. The anti-OA response in recipient mice was dependent on the presence of fetal calf serum (FCS) in the medium in which the cells were suspended. Primed spleen cells suspended in serum-free medium containing lipopolysaccharide (LPS) also transferred the anti-OA response without requiring antigenic challenge of the recipients. No antigenic similarity could be shown to exist between OA, FCS, and LPS in either PCA or HA assay. The transfer of the response in the absence of exogenous antigen evidently depended on a mitogenic stimulus to the primed spleen cells. However, anti-OA antibody injected into irradiated recipients immediately before transfer of primed cells suppressed the production of both IgE and hemagglutinating antibodies to OA in the recipient. This implied that antigen was present in cell-bound form on primed cells. Possible application of this model to the study of the regulation of IgE production in an ongoing response is discussed.
Two-Gene Control of the Expression of Murine Ia Antigens
Authors: Jones Patricia P. , Murphy Donal B. , McDevitt Hugh O.
Published: December 2022

Two-dimensional polyacrylamide gel electrophoresis of Ia antigens obtained from 35S-methionine-labeled lymphocytes has revealed that the expression on the cell surface of an antigen coded for by a locus in the I-A subregion may be controlled by an I-E subregion product. Three distinct polypeptide chains (Aa, Ab, and Ae) coded for by loci in I-A have been identified in gels of immunoprecipitated Ia antigens; one of these (Ae) is coprecipitated with an I-E antigen by anti-I-E antibodies and probably is complexed to the I-E chain in the cell.

Mice of haplotypes which do not have serologically-detectable I-E antigens (such as b and s haplotypes) appear not to synthesize and I-E product. These mice also do not express Ae on their cell surfaces, although precursors of surface Ae can be detected in the cytoplasm. Some haplotypes, however, contain a locus mapping between Ia.4 in I-J and H-2D which can complement I-A in either the cis or trans position to allow the cell surface expression of Ae.
Structural Characterization of Murine Ia Alloantigens
Authors: Cook Richard , Capra J. Donald , Vitetta Ellen S. , Uhr Jonathan W.
Published: December 2022

The murine Ia alloantigens encoded by the I-A and I-E/C subregions were examined by SDS-polyacrylamide gel electrophoresis and microsequence analysis. By sensitive double label experiments (14C and 3H), the alpha (α) and beta (β) polypeptides of the A and E/C subregions were analyzed for molecular weight (mw) variation. The α and β chains of the A subregion (k, b, and d haplotypes) electrophoresed at 34,000 and 26,000 daltons, respectively. Haplotype associated differences in mw were detected for the E/C subregion products — k and r haplotype subunit mw were 34,000 (α) and 28,000 (β) daltons, whereas the α and β chains of p and d haplotypes were 31,000 and 29,000 daltons, respectively.

The α and β chains of both the A and E/C antigens do not exist on the cell surface as disulfide bonded α plus β forms.
Anti-Thy-1.1 Response: Possible Relationship Between Ir Control and Antigenic Competition Involving H-2 Antigens
Authors: Zaleski M. , Gorzynski T. , Klein J.
Published: December 2022

Mice were immunized with Thy-1.1 disparate thymocytes obtained from either H-2 compatible or H-2 incompatible donors. The magnitude of the ensuing anti-Thy-1.1 response was measured by the number of PFC in spleens of experimental animals. H-2 compatible thymocytes almost always elicited a significantly higher response than that induced by H-2 incompatible thymocytes. Incompatibility between donor and recipient limited to either the K or D region alone resulted in a response comparable to that produced by thymocytes incompatible over the entire H-2 complex. It seems that products of incompatible H-2 alleles inhibit the response to Thy-1.1 alloantigen. This is indicated by findings that: 1) immunization with a mixture of H-2 compatible and incompatible, or with H-2 heterozygous thymocytes elicited a lower anti-Thy-1.1 response than that produced by H-2 compatible cells alone, 2) preimmunization with H-2 incompatible cells inhibited the subsequent response to H-2 compatible thymocytes, 3) tolerance to antigens of an incompatible H-2 haplotype abrogated the adverse effect of these antigens on anti-Thy-1.1 response.
Role of IJ Region Determinants in Neonatal Tolerance Induction to K and D Region Alloantigens
Authors: Streilein J. W. , Klein J.
Published: December 2022

Tolerance induction to isolated regional H-2 alloantigenic differences was studied by inoculating neonatal animals with lymphohematopoietic cells from F1 hybrids derived from H-2 congenic pairs, intra-H-2 recombinants. H-2 regions varied considerably in the tolerogenicity of their alloantigenic products, especially when host animals were derived from B10 and B6 genetic backgrounds. Isolated K and D region disparities, especially those resulting from mutation, were highly resistant to tolerance induction, while I region differences alone were highly tolerogenic; ease of tolerance induction across the entire H-2 complex was intermediate between these extremes. Using appropriate recombinants, poorly tolerogenic D and K region determinants, when presented conjointly with IJ region disparity in the tolerance conferring inoculum, proved to be highly specific, not only for the individual antigenic determinants, but apparently also for the composite antigenicity of the tolerance conferring inoculum: tolerance of D alloantigens was restricted to their original IJ region context, i.e. skin grafts from third party donors expressing the tolerated D antigen conjointly with an IJ region determinant unexpressed in the tolerizing inoculum were rejected.
Ir Gene Regulation of Accessory Cell Function
Authors: Singer Alfred , Cowing Carol C. , Dickler Howard B. , Hodes Richard J.
Published: December 2022

The possibility was investigated that Ir genes regulate the function of cells other than T or B cells in the primary IgM PFC responses to the synthetic antigens TNP-(T,G)-A--L and TNP-(H,G)-A--L. The primary PFC responses of (Responder × Nonresponder)F1 spleen cells to both antigens were abrogated by G-10 Sephadex passage and restored by the addition of spleen adherent cells. The cell type in the spleen adherent cell population active in reconstituting the responses to TNP-(T,G)-A--L and TNP-(H,G)-A--L was a non-T, non-B, radiation resistant, glass adherent spleen cell. The abrogated responses of G-10 Sephadex passed (Responder × Nonresponder)F1 spleen cells to each antigen were reconstituted by spleen adherent cells from only strains which were responders to that antigen. All the spleen adherent cells tested restored the non-Ir gene controlled response to a third antigen, TNP-KLH.
Genetic Restriction of T-Macrophage Interactions: Is It Induced by Priming or Present Prior to Priming?
Authors: Erb Peter , Feldmann Marc
Published: December 2022

There is now ample evidence that T-macrophage interactions in the induction of various immune responses are genetically restricted. We found that the in vitro generation of T helper cells with soluble antigens requires I-A compatible macrophages, or GRF (genetically related macrophage factor) a complex of Ia products and antigenic fragments. However, it is unclear whether genetic restriction of T-macrophage interactions is imposed by priming or already expressed prior to priming. Using F1 T cells and parental macrophages we were able to generate helper cells that cooperated preferentially with B cells and macrophages of the parental haplotype used for initial priming, indicating that genetic restriction may be induced by priming. However, results obtained by using T cells from chimeric mice instead of F1 hybrids suggested that the restriction of T-macrophage interaction may be already present prior to priming. This conclusion is supported by absorption experiments — unprimed splenic T cells or thymocytes preferentially adsorb I-A compatible GRF.
Evidence for an Association Between TNP-Conjugated Macrophage Membrane Components and IA Antigens
Authors: Thomas David W. , Shevach Ethan M.
Published: December 2022

The effects of anti-TNP and anti-Ia sera on the ability of TNP-modified macrophages to stimulate TNP-specific T cell proliferative responses in vitro were examined. T cell responses to fresh TNP-modified macrophages were dramatically reduced in the presence of anti-TNP serum. By contrast, anti-TNP had no effect on the stimulatory capacity of TNP-modified macrophages that had been cultured overnight (aged) prior to addition to responder cells. However, the addition of anti-Ia serum inhibited the proliferative response to both aged and fresh TNP-modified macrophages. Moreover, there was no requirement that anti-Ia serum be present continuously, since brief anti-Ia treatment of TNP-modified macrophages (20% anti-Ia, 60 min at 37°C) also reduced their stimulatory capacity. The inhibition of stimulation by TNP-modified F1 macrophages briefly treated with anti-Ia directed against one or the other parents was haplotype-specific indicating that anti-Ia did not non-specifically interfere with macrophage presentation of TNP.
The Genetic Restrictions on Macrophage-T Cell Interaction Lead to the Genetic Restrictions of T Cell-B Cell Interaction
Authors: Yamashita U. , Shevach E. M.
Published: December 2022

Although the histocompatibility restriction for the interaction of antigen-pulsed macrophages and proliferative T cells have been confirmed, there exists considerable controversy on the histocompatibility restrictions for macrophage-helper T cell interaction and for T cell-B cell interaction. In order to study the histocompatibility restriction between macrophages and helper T cells, carrier primed guinea pig T cells were positively selected in vitro with antigen-pulsed macrophages for 7 days and the selected T cells were then mixed with hapten-primed B cells and stimulated with antigen in a modified Mishell-Dutton system. (2 × 13)F1 T cells selected with OVA-pulsed F1 macrophages cooperated with B cells of both parents. However, F1 T cells selected with strain 2 macrophages cooperated with strain 2 B cells, but not strain 13, while F1 T cells selected with strain 13 macrophages cooperated with strain 13 B cells, but not strain 2.
The Nature of MHC Restrictions in T-B Cooperation
Authors: Waldmann H. , Pope H. , Brent L.
Published: December 2022

The I region of the mouse major histocompatibility complex plays an important role in controlling the interactions between T helper cells and B cells in response to antigen. For example, MHC incompatible T cells from BALB/c mice cooperate well with BALB/c B cells but poorly with CBA B cells in antibody formation. Data will be presented on helper T cell function in the PFC response to TNP-KLH in four sorts of mice, namely: a) double bone marrow chimeras (e.g. C3H + BALB/c stem cells → irradiated (C3H × BALB/c) F1 recipients), b) single chimeras (e.g. C3H or BALB/c stem cells into (C3H × BALB/c) F1 recipients), c) F1 → Parental chimeras (e.g. (C3H × BALB/c) F1 stem cells into C3H or BALB/c recipients), d) CBA mice rendered neonatally tolerant to BALB/c spleen cells.

These studies suggest that the following factors determine optimal interaction between MHC incompatible T and B cells.
Biological Significance of Protein-Defined and Carbohydrate-Defined Ia Antigens
Authors: Parish Christopher R.
Published: December 2022

Recent studies suggest that the I-region may control two separate classes of antigens, one class consisting of protein-defined antigenic determinants and the other consisting of carbohydrate-defined specificities. It was important to determine the contribution of these two classes of Ia antigens to various I-region controlled immune phenomena. Initially, the nature of the Ia antigens carried by antigen-specific helper factors secreted by T cells was investigated. It was found that both GAT-specific and TGAL-specific helper factors from CBA/H mice (H-2k) were retained and eluted from immunoabsorbents which contained rabbit antibodies directed exclusively against the carbohydrate-defined Ia antigens (anti-Iak). Mouse anti-Iak immunoabsorbents which carried antibodies against both the carbohydrate-defined and protein-defined Ia antigens also removed both helper factors. However, these immunoabsorbents were unable to bind helper factor when they were pretreated with a dialyzable material in normal (H-2k) mouse serum which is known to neutralise antibodies against the carbohydrate-defined Ia antigens.
I-C Subregion Determinants on MLR Suppressor Factor
Authors: Rich Susan Solliday , David Chella , Rich Robert R.
Published: December 2022

MLR suppressor factor (MLR-SF) optimally suppresses MLR responses only of responder cells which are compatible in the I-C subregion with cells producing the factor. Therefore, presence of MHC-encoded determinants on MLR-SF has been investigated, with particular interest in I-C, as well as the possible role of I-J determinants. Suppressor activity of supernates of alloantigen-activated spleen cells was tested after passage over solid immunoadsorbants prepared with various anti-H-2 subregion sera. Immunoadsorbants with specificity towards K and D region products were ineffective while several preparations with I region specificity removed suppressor activity from factors of the appropriate haplotype. The single subregion specificity common to all adsorbing preparations was I-C. Recently, ambiguity has developed with regard to genetic resolution of I-C and adjacent subregions, including difficulties of serologic differentiation of I-C products of the k and d haplotypes. Antisera prepared in I-Ck/I-Cd disparate strain combinations have identified T cell restricted determinants which are distinct between these haplotypes.
Cross Reactive Idiotypic Determinants on (T,G)-A--L Specific Antibodies of High and Low Responder Mice
Authors: Mozes Edna , Lifshitz Ruth , Schwartz Michal
Published: December 2022

Mice possessing the H-2b haplotype produce high antibody titers when immunized with (T,G)-A--L, whereas mice possessing the H-2k haplotype are low responders to this immunogen. Following immunization with either (Phe,G)-A--L or (T,G)-A--L complexed with methylated bovine serum albumin (MBSA), low responder as well as high responder mice produce antibodies which can be titrated with (T,G)-A--L. The variable regions of antibodies produced against the above immunogens were compared utilizing guinea pig sera against (T,G)-A--L idiotypes of C3H.SW (H-2b, Ig-1a) mice, which appear to be allotype linked. (T,G)-A--L specific antibodies were isolated from sera of H-2b (Ig-1a) and H-2k (Ig-1a) mice immunized with (Phe,G)-A--L. These antibodies inhibited 50% of the 125I-idiotype to the anti-idiotypes. In contrast, a population of antibodies isolated from the (Phe,G)-A--L specific antisera which bound only 125I-modified (Phe,G)-A--L did not react with the anti idiotypes.
Enhanced Susceptibility of Virus-Infected Fibroblasts to Cytostasis Mediated by Peritoneal Exudate Cells
Authors: Goldman Rachel , Hogg Nancy
Published: December 2022

Murine (BALB/c) starch-induced adherent peritoneal exudate cells (PEC) exert a cytostatic effect on various tissue culture cell lines. The cytostatic activity resides in the adherent cell fraction of the PEC that, after 24 hr in culture, is devoid of T and B lymphocytes. No significant difference in the degree of inhibition of incorporation of DNA precursors upon interaction with PEC could be detected with transformed cell lines, virus-infected non-transformed cell lines, and their normal counterparts as targets.

Recently explanted embryo fibroblasts are relatively resistant to PEC-mediated cytostasis. Upon in vitro transformation with the murine sarcoma virus murine leukemia virus mixture (MuSV/MuLV), the targets become more susceptible to PEC-mediated cytostasis. Infection with the nontransforming virus MuLV leads to a similar increase in susceptibility. The same pattern of differential susceptibility was obtained with embryo fibroblasts from four different strains of mice, indicating that there is no requirement for H-2 compatibility between effector and target cells. No PEC-specific 51Cr release could be detected during the 24 hr of effector-target cell interaction, indicating that the effect involves primarily cytostasis.

Embryo fibroblasts infected with Sendai virus or influenza virus were also remarkably sensitized toward PEC-mediated cytostasis. The fact that inactivated Sendai virus is as active in sensitizing the targets as the active virus suggests that primary membrane reorganization events that follow virus adsorption may be a sufficient perturbation leading to enhanced susceptibility.

Rat and hamster embryo fibroblasts show the same pattern of enhanced susceptibility upon interaction with inactive Sendai virus. Judged from indirect radioimmunoassay of adsorption of anti-type-C viral antigen sera, BALB/c embryo fibroblasts did not exhibit surface type-C virus specific antigens, nor did infection with the Sendai or influenza viruses induce activation of endogenous type-C viruses. The above suggests that transformation and a major alteration in the growth pattern of cells are not a prerequisite for the enhanced susceptibility of embryo fibroblasts to PEC-mediated cytostasis; rather, that membrane perturbations introduced by nontransforming unrelated viruses provide a sufficient change rendering the cells more prone to cytostasis.
Immunochromatography on Insolubilized Antibodies of Very Low Affinity: Application to Immunoadsorbence of Bovine α-Fetoprotein
Authors: Ruoslahti Erkki
Published: December 2022

Cross-reactive antibodies were utilized to prepare immunoadsorbents possessing a very low affinity to bovine α-fetoprotein (AFP). A goat anti-human AFP serum cross-reactive with bovine AFP was first depleted of antibodies reactive with bovine AFP in immunodiffusion. The remaining antibodies from this serum and γ-globulin from a sheep antiserum against rabbit AFP, without prior absorption, were coupled to Sepharose. Chromatography of fetal calf serum on these adsorbents resulted in retardation of bovine AFP relative to other proteins. A major part of the AFP eluted from the columns with phosphate-buffered saline. The rest eluted as a sharp peak with a small quantity of 4 or 6 M urea.

The elution of AFP with the initial column buffer has made it possible to prepare pure AFP that has not been subjected to the chaotropic elution buffers usually employed in affinity chromatography. Elimination of the washing step and the ease of elution has allowed purification of gram amounts of AFP. The fact that immunoadsorbents prepared from antibodies with no detectable reactivity in immunodiffusion still caused delayed elution in chromatography suggests that this procedure may be useful in search of proteins cross-reactive with a known protein.
Bursectomy in ovo Blocks the Generation of Immunoglobulin Diversity
Authors: Huang Henry V. , Dreyer William J.
Published: December 2022

Immunoglobulins made by chickens bursectomized in ovo on day 11 of incubation were studied by two-dimensional gel electrophoresis. All such bursectomized chickens have limited diversity in their immunoglobulin molecules. A range of different degrees of diversity restriction was found in individual bursectomized chickens. The limited immunoglobulin diversity was stable for at least 20 weeks in that the same spots were found over this time span. Bursectomized chickens that responded to repeated antigen challenge had sheep erythrocyte- and dinitrophenol-specific antibodies of limited diversity. Different bursectomized responders made almost identical antigen-specific antibodies. The results were interpreted as indicating that individual bursectomized birds had been blocked at different points during the developmental generation of immunoglobulin diversity, and that the genetic mechanisms that underlie the developmental diversification of clones of B cells lead to a sequential generation of very similar antibody molecules in different embryos.
Lymphocyte Transformation Induced by Chemical Modification of Membrane Components
Authors: Beyer Carl F. , Bowers William E.
Published: December 2022

Neuraminidase treatment of responder cells in indirect periodate or indirect neuraminidase-galactose oxidase (NGO) stimulation of rat lymph node cells (LNC) led to proliferative responses that were substantially greater and faster than those of controls. The indirect responses followed bicellular kinetics, i.e., the number of cells that initially responded was a function of the product of the number of stimulator cells and the number of responder cells in the cultures.

Indirect NGO stimulation between allogeneic stimulators and responders was very similar to that between syngeneic cells, and neuraminidase treatment of responders had similar effects in both cases. The indirect mitogenic response between allogeneic cells completely abrogated the expected mixed lymphocyte reaction (MLR).

Neuraminidase treatment of stimulators had little effect on the kinetics or magnitude of the rat one-way MLR. Treatment of responders, however, led to a faster and stronger response as compared with the normal MLR response. Treatment of both stimulators and responders was essentially the same as treatment of responders alone. These results, and those for indirect stimulation, suggest that the ability of responder cells to receive a mitogenic signal from stimulator cells is modulated by the amount of sialic acid on the responder cell surface.

Although more blast cells were produced in indirect NGO cultures with neuraminidase-treated responders, the cytotoxic activity of these cultures was not enhanced over that produced in normal indirect stimulation. In fact, when referred to the extent of cellular proliferation in each culture, the cytotoxic activity produced by indirect stimulation was higher than that produced either by direct NGO stimulation or by indirect stimulation with neuraminidase-treated responders. Thus indirect NGO stimulation selectively activates cytotoxic cells. The additional cells that are activated by direct NGO stimulation, or after neuraminidase treatment of indirect responders, do not appear to be cytotoxic.