bluuebunny/crossref_metadata_2025 huggingface dataset:
T lymphocytes of young and aged rats. II. Functional defects and the role of interleukin-2.
Authors: Gilman S C , Rosenberg J S , Feldman J D
Published: December 2022

Splenic T lymphocytes of aged Lewis rats respond to Con A and PHA with diminished 3H-TdR uptake compared with splenic T lymphocytes of young Lewis rats. After immunization with allogeneic tumor cells, uptake of 3H-TdR in mixed lymphocyte-tumor cultures and T cell cytotoxicity against tumor target cells are significantly lower with spleen cells of aged rats compared with those of young rats. The culture of spleen cells of aged rats with Con A results in a diminished conversion of Ia-positive T cells from Ia-negative precursors compared with similar cultures of spleen cells of young rats. Spleen cells of both young and aged rats produce high amounts of IL-2 in response to Con A stimulation. "Old" T cells, however, bind relatively little IL-2, do not utilize it in culture, and do not respond to exogenous IL-2 with enhanced 3H-TdR uptake as do "young" T cells. In allogeneic MLTC, "old" T lymphocytes produce little IL-2 compared with "young" cells, and both "young" and "old" cells respond to exogenous IL-2 with enhanced 3H-TdR uptake and increased cytotoxic activity. The data suggest defects in the synthesis and/or recognition of IL-2 as well as defects in the regulation of Ia antigen expression may be responsible, in part, for the reduced T cell function in aged animals.
The requirement for esterase activation in T cell replacing factor (TRF)-induced IgG production in a human B blastoid cell line.
Authors: Miki Y , Kishi H , Muraguchi A , Kishimoto S , Yamamura Y , Kishimoto T
Published: December 2022

Involvement of stimulus-activatable serine esterase in the process of TRF-induced IgG production in a human B blastoid cell line (CESS) was demonstrated. Addition of DFP or PMSF (1 X 10(-3) M to 1 X 10(-5) M) showed a dose-dependent inhibitory effect on TRF-induced IgG induction in CESS cells. Other organophosphorous inhibitors, such as p-NPEPP and PEPP, were also effective, but a nonphosphorylating analogue of DFP, DMP, was not inhibitory. Simultaneous addition of DFP with TRF showed the inhibitory effect, and the inhibition was irreversible. However, pretreatment of the cells with DFP did not abrogate TRF-induced IgG production. In synchronized cells, signals provided by TRF could be transduced only when cells were in the G1-phase, and DFP showed an inhibitory effect only when it was added to cells in the G1-phase with TRF. One amino acid derivative (AAME) partially inhibited TRF-induced IgG production.
Studies on induction and effector functions of concanavalin A-induced suppressor cells that limit TCGF production.
Authors: Gullberg M , Larsson E L
Published: December 2022

The cellular mechanisms regulating TCGF production in Con A-stimulated cultures have been investigated. Normal spleen cells, activated by Con A for 24 hr, develop suppressive cells that inhibit de novo production of TCGF by fresh spleen cells. Effector cells mediating suppression are nonadherent, radioresistant, Lyt-2-positive T cells. The induction of suppressor cells is radiosensitive and it requires 18 hr. The kinetics of suppressor cell induction parallels very closely the termination of TCGF production in situ, suggesting the major importance of this mechanism in the control of TCGF production. Reculture of 24-hr Con A-activated cells in the absence of Con A for 24 to 72 hr results in a gradual loss of suppressive activity that can be recalled by readdition of Con A with the same kinetics found in fresh spleen cells. In addition, de novo production of TCGF is readily induced in such cultures upon restimulation with Con A, demonstrating that abrogation of TCGF-production in primary cultures is due to suppression and not to lectin-dependent killing of the TCGF-producing T cells. Measurements of suppressive activity and lectin-dependent cytotoxicity, in various populations of Con A-activated spleen cells, further distinguish these activities. Finally, the reduced suppressive activity of Con A-activated cells after expansion in TCGF excludes that suppressor cells act by absorption or removal of TCGF produced at normal rates. Direct, reversible suppression of TCGF-producing cells by T lymphocytes appears, therefore, to constitute a major mechanism by which cytotoxic T cell responses are regulated.
Suppressor-target interaction in alloantigen induced responses: induction of a second cell in the suppressive pathway.
Authors: Beckwith M , Rich S S
Published: December 2022

The sequence of events involved in MLR-TsF-induced suppression of the mixed lymphocyte response has been investigated. It is demonstrated in this report that MLR-TsF, which is secreted by predominantly Ly-2+ alloantigen-primed splenic T cells, carries out its suppressive function through the active recruitment of an intermediary suppressor cell. Substantial suppression of the proliferative response is observed when unprimed BALB/c splenocytes are cultured with MLR-TsF and then added as irradiated regulator cells to MLR. This inductive process is dependent on factor concentration, requires a 48-hr exposure of cells to MLR-TsF, and occurs in the absence of added priming strain alloantigen. The target cell involved in this factor-cell interaction is characterized as a nonadherent, Ly-1+2+ T cell resistant to complement-dependent cytotoxicity with a monoclonal anti-Thy-1.2 antibody. In addition, second-order suppression does not appear to operate in an antigen-specific manner. These studies indicate that MLR suppression is a complex process involving a series of interactions between cellular components and their soluble regulatory products.
Differences in the migration of B and T lymphocytes: organ-selective localization in vivo and the role of lymphocyte-endothelial cell recognition.
Authors: Stevens S K , Weissman I L , Butcher E C
Published: December 2022

The migration of B and T lymphocytes in the mouse has been studied by using 1) short-term in vivo homing studies, and 2) an in vitro assay of lymphocyte binding to specialized lymphoid organ venules (post-capillary, high endothelial venules (HEV)) in frozen sections of lymph nodes and Peyer's patches. The homing characteristics of B and T cell populations are largely independent of their organ of origin. B cells from any source distribute preferentially to Peyer's patches, whereas T cells home preferentially to peripheral lymph nodes. This organ specificity of migration appears to be determined at the site of lymphocyte exit from the blood by selective recognition of organ-specific determinants on the endothelial cells of HEV. In addition, the in vivo tendency of B cells to migrate preferentially to the spleen, and of T cells to localize better in lymph nodes is confirmed. The results indicate that, in a hypothetical situation in which an equal number of B and T lymphocytes localized in peripheral lymph nodes (or bound in vitro to peripheral node HEV), there would be about 2.5 B cells for every T cell in the mesenteric node, four to six B cells per T cell in Peyer's patches, and seven to nine B cells per T cell in the spleen. Comparison of these homing preferences with the distribution of B and T lymphocyte populations in situ suggests that selective lymphocyte migration may help determine the proportions of functionally distinct lymphocyte classes in particular lymphoid organs or sites of chronic inflammation, and thus may serve to influence the character of local immune responses.
Mechanism of enhanced complement-dependent cytotoxicity of papain-treated lymphocytes: evidence for increased stability of classical pathway C3 convertase.
Authors: Lublin D M , Grumet F C
Published: December 2022

The mechanisms of enhanced antibody-mediated, complement-dependent cytotoxicity caused by proteolytic treatment of cells was studied in a model system based on HLA microlymphocytotoxicity methods. In this model, papain treatment of lymphocytes resulted in a) no change in antibody binding, b) a slight decrease in initial binding of C4, c) a marked increase in stability of cell-bound C4b, resulting in d) an increase of cell-bound C3, and e) no increase in lytic efficiency of the C5b-9 membrane attack complex. We conclude that the most important step in papain enhancement of lysis of lymphocytes is an increased stability of cell-bound C4b, possibly through decreased surface binding of C4-bp. This mechanism may be relevant to the pathologically increased lysis of cells occurring in patients with hereditary erythroblastic multinuclearity with a positive acidified-serum test (HEM-PAS).
Microinjection of macromolecules into normal murine lymphocytes by cell fusion technique. I. Quantitative microinjection of antibodies into normal splenic lymphocytes.
Authors: Ohara J , Watanabe T
Published: December 2022

Human erythrocyte ghosts loaded with various kinds of protein molecules were fused with mouse splenic lymphocytes by means of polyethylene glycol supplemented with poly-L-arginine and dimethylsulfoxide. This fusion method made quantitative microinjection of IgG and other proteins into intact lymphocytes possible. The injection itself did not alter cell viability, and lymphocytes given protein molecules retained intact response activity when they were stimulated with mitogens. Rabbit anticyclic AMP was purified by affinity chromatography and injected into lymphocytes. Antibody activity in the cell lysates was measured by using 125I-labeled cyclic AMP as an antigen, and it was shown that antibody molecules were quantitatively injected and immunologically active in the cells. Antigen binding activity of anti-cyclic AMP antibodies in the nonstimulated lymphocytes was stable and intact even 24 hr after microinjection, whereas the activity rapidly decreased in mitogen-stimulated lymphocytes, indicating that some immunologic or enzymatic mechanisms for inactivating antibodies were induced in mitogen-stimulated cells. Furthermore, microinjection of anti-cyclic AMP markedly enhanced the proliferative responses of lymphocytes to mitogens such as Con A or LPS and reversed the effect of a known elevator of intracellular cyclic AMP. These observations have implications for the role of cyclic AMP in early lymphocyte activation events.
The purification and properties of human T cell growth factor.
Authors: Mier J W , Gallo R C
Published: December 2022

Human T cell growth factor (TCGF) has been purified more than 800-fold from serum-free lymphocyte conditioned media by utilizing ion exchange chromatography with DEAE-Sepharose, gel filtration with Ultrogel AcA54, and preparative SDS-polyacrylamide gel electrophoresis. This mitogenic protein, which is released by T cells into the incubating media after exposure to a lectin (PHA), has a m.w. of 13,000 as determined by SDS-PAGE and 20,000 to 25,000 on gel filtration and has an isoelectric point of 6.8. The material extracted from acrylamide gels is a single band (or two nearly superimposed bands) when rerun on analytical SDS gels. Partially purified material is unstable even at -70 degrees C and requires the addition of bovine serum albumin or polyethylene glycol to maintain biologic activity. It is sensitive to proteolytic digestion but resistant to nucleases and thiol-reducing agents such as dithiothreitol. Human TCGF can be reversibly denatured with urea or SDS. Material extracted from acrylamide gels has been shown to sustain T lymphoblasts in tissue culture. In contrast to lectins, certain antigens, and crude lymphocyte conditioned media, purified TCGF does not initiate lymphocyte blastogenesis but is a highly selective mitogen for T cells previously activated by exposure to lectins or antigens.
Anti-RNP monoclonal antibodies derived from a mouse strain with lupus-like autoimmunity.
Authors: Billings P B , Allen R W , Jensen F C , Hoch S O
Published: December 2022

systemic lupus erythematosus (SLE) and related rheumatic and connective-tissue diseases are often associated with the production of antibodies directed against a variety of specific cellular components. Recent evidence indicates that two such autoantigens, the Sm and RNP antigens recognized by SLE sera, exist in small ribonucleoprotein complexes found in the nuclei of higher eukaryotes. Studies of the structure and function of these autoantigenic particles with human sera used as probes have been limited because of the multiplicity of autoantibodies often found in an individual serum. Through this communication, we report that MRL/Mp-+/+ (MRL/n) mice, which spontaneously develop a disease exhibiting many of the characteristics of human SLE, possess anti-RNP antibodies in addition to anti-Sm and anti-DNA as previously reported. Spleen cells from one such autoimmune mouse were used to produce a stable hybridoma secreting antibodies that react simultaneously with a protein of Mr 40,000 and a doublet of approximately 70,000, a pattern of reactivity identical to and characteristic of human SLE anti-RNP autoantibodies.
Sr++-induced inhibition of human natural killer (NK) cell-mediated cytotoxicity.
Authors: Neighbour P A , Huberman H S
Published: December 2022

The cell believed to be responsible for human spontaneous cytotoxicity or natural killing (NK) has been described recently as a large granular lymphocyte (LGL). Because the intracytoplasmic granules of LGL appear to be peculiar to the subpopulation of mononuclear cells containing NK effectors, we have investigated their possible requirement in the cytolytic process. Experiments reported here show that Sr++, a cation known to cause degranulation of granulocytes, caused a dose- and time-dependent inhibition of the NK activity of human peripheral blood mononuclear cells. A single-cell NK assay showed that the number of cytotoxic cells was not significantly reduced, but that Sr++ inhibited NK by preventing individual effector cells from engaging in multiple lethal hits. Moreover, the inhibition of NK appeared to be reversible; culture in vitro of Sr++-treated PBMC resulted in the eventual return of cytolytic activity in these cells. Augmentation of NK by interferon (IFN) was similarly blocked by Sr++. In addition, Sr++ also inhibited the NK activity of IFN-prestimulated effector cells and, as before, NK function returned during subsequent culture. These results indicated that the return of NK function after Sr++ treatment was the result of reactivation of cytotoxic cells rather than recruitment of new effectors. We have shown in other studies that Sr++ degranulates LGL, and the data presented here provide support for the hypothesis that the intracytoplasmic granules of LGL are required for NK-mediated cytotoxicity.
Monoclonal antibody to Mullerian inhibiting substance.
Authors: Mudgett-Hunter M , Budzik G P , Sullivan M , Donahoe P K
Published: December 2022

Mullerian Inhibiting Substance (MIS) is a testicular product that causes the regression of the Mullerian duct in the developing male embryo. Antibody specific for MIS would facilitate the purification and study of this "hormone," but because of its impure status, traditional polyclonal antisera specific for MIS would be untenable. The requisite specificity, however, might be obtained by the technique of somatic fusion, regardless of the purity of the immunizing antigen. This paper describes the production of 2 monoclonal antibodies specific for MIS by the technique of somatic cell fusion. Spleen cells from mice immunized with an impure preparation of MIS were fused with myeloma cell line NS1. Culture media from the resulting hybridoma cell lines were screened for anti-MIS antibody by a sensitive RIA. Specificity for MIS was demonstrated by the adsorption of biologically active MIS on an affinity column prepared from monoclonal anti-MIS antibody. MIS activity as assessed by an organ culture assay was subsequently recovered from the affinity column in the fraction eluted with MH4SCN. Using the RIA, monoclonal anti-MIS antibody was also shown to compete favorably with a variety of potentially cross-reactive proteins.
Anti-immunoglobulin stimulation of murine lymphocytes. VII. Identification of an age-dependent change in accessory cell function.
Authors: Scribner D J , Moorhead J W
Published: December 2022

Rabbit anti-mouse immunoglobulin-induced proliferation of murine B cells requires the participation of a macrophage- (M phi)like accessory cell. This cell is found in the spleen of aged mice, the B cells of which respond to stimulation by intact anti-Ig molecules. M phi characteristics of this cell include resistance to treatment with either anti-theta or anti-Ig plus complement, adherence to plastic, and resistance to radiation. Evidence was obtained that indicates that the age-dependent anti-Ig responsiveness of B cells involves, in part, a temporal change in the splenic M phi population. The ability of these accessory cells to facilitate anti-Ig responsiveness by normally unresponsive B cells was shown both in vivo and in vitro. Reconstitution of lethally irradiated aged, but not young, mice with spleen cells from unresponsive young mice resulted in the appearance of anti-Ig responsive B cells within 30 days. Similarly, coculture of unresponsive spleen cells with irradiated splenic adherent cells from aged, but not young, mice provided accessory cell activity for anti-Ig responses. Additional experiments showed that accessory cells were also present in bone marrow and the peritoneal cavity but were absent in lymph node and thymus. Addition of 2-mercaptoethanol to the culture medium functionally replaced the requirement for accessory cells and was also found to alter the response pattern of the B cells.
Evidence for the presence of suppressor T lymphocytes in animals treated with cyclosporin A.
Authors: Wang B S , Heacock E H , Chang-Xue Z , Tilney N L , Strom T B , Mannick J A
Published: December 2022

Mice sensitized with alloantigens and treated with cyclosporin A (CsA) were incapable of generating antigen-specific cytolytic lymphocytes (CL). Lymphocytes from these CsA-treated animals could not be reactivated upon exposure to the same alloantigens in mixed lymphocyte culture (MLC), whereas their response to a third-party antigen remained intact, suggesting a long-lasting and specific effect of CsA. After being irradiated, these lymphocytes from CsA-treated animals were added to normal MLC and were shown to prevent normal lymphocytes from becoming cytolytic in a dose-dependent and antigen-nonspecific fashion. These suppressor cells were not detected in mice receiving CsA only, indicating that CsA did not induce but rather permitted the expression of suppressor cells possibly generated by allosensitization. The suppressor cells appeared to be T lymphocytes, because treatment with anti-Thy-1.2 antibody and C abrogated their suppressive activity. The present results suggest that activation and/or sparing of suppressor cells by CsA may account for the long-lasting unresponsiveness seen in CsA-treated animals.
Adoptive transfer of modulation of granuloma formation and hepatosplenic disease in murine schistosomiasis japonica by serum from chronically infected animals.
Authors: Olds G R , Olveda R , Tracy J W , Mahmoud A A
Published: December 2022

The course of disease due to Schistosoma japonicum in C57BL/6J mice was examined. Animals infected with 25 cercariae of the Philippine strain of S. japonicum for 9 wk develop a peak portal hypertension of 13 +/- 0.6 cm H2O and mean hepatic granuloma area of 459 X 10(3) micrometers 2. By 30 wk, disease manifestations were reduced; mean portal pressure was 7 +/- 0.5 cm H2O and mean hepatic granuloma area was 12 X 10(3) micrometers 2. Adoptive transfer of serum but not 1 X 10(8) spleen or 4 X 10(7) lymph node cells from chronically infected mice resulted in lowering portal pressure and decreasing granuloma size in recipients. Granuloma formation in the sensitized lung model was also suppressed by serum from chronically infected mice. The immunosuppressive component of this serum was localized in the IgG1 fraction.
Interactions of bacterial lipopolysaccharide with acute-phase rabbit serum and isolation of two forms of rabbit serum amyloid A.
Authors: Tobias P S , McAdam K P , Ulevitch R J
Published: December 2022

Mixing lipopolysaccharide (LPS, Salmonella minnesota R595) with acute-phase rabbit serum (APRS) results in the formation of two types of complexes. The two complexes are separable from each other and from LPS by CsCl density gradient ultracentrifugation. LPS alone had density 1.38 g/cm3, complex 1.3 had density 1.3 +/- 0.02 g/cm3, and complex 1.2 had density less than 1.20 g/cm3. At 1 to 10 micrograms LPS/ml APRS, up to 23 micrograms of LPS could be found in complex 1.3, with the remainder of the LPS in complex 1.2. The capacity of complex 1.2 for LPS was 500 to 600 micrograms LPS/ml APRS. The ability of rabbit serum to form complex 1.3 rose to a maximum in 24 hr post-acute-phase induction. The two complexes were purified by equilibrium density gradient ultracentrifugation and immunoaffinity chromatography using antibodies to LPS and subjected to SDS-PAGE. Complex 1.3 had major peptides after reduction of 91, 64, 61 and 50 kilodaltons. The 61-kilodalton peptide is probably rabbit serum albumin; the others are unidentified. Complex 1.2 had major peptides after reduction whose mobility corresponded to apolipoprotein A-I and serum amyloid A. Complex 1.2 thus seems to be analogous to the LPS-high-density-lipoprotein complexes that form in normal serum except that the latter complexes do not contain serum amyloid A. Rabbit serum amyloid A (SAA) was purified by CsCl equilibrium density gradient ultracentrifugation, gel filtration, and ion-exchange chromatography into two forms, SAA-1 and SAA-2. The two forms of SAA have very similar amino acid compositions and m.w. (SAA-1, 11,526 daltons; SAA-2, 11,469 daltons). They differ primarily in their isoelectric points, 6.57 and 6.27 for SAA-1 and SAA-2, respectively. The complexes 1.2 from several individual rabbits after acute-phase induction were studied by isoelectric focusing. Seven of 10 rabbits showed both forms of SAA; three showed a single form. Two rabbits showing SAA-1 on the first acute-phase induction were reinduced; one rabbit again gave SAA-1 and the other gave SAA-2. These results suggest that SAA and perhaps other acute-phase reactants may modulate the biologic activity of LPS.
Immunogenetics of BCG-induced anergy in mice: control by genes linked to the Igh complex.
Authors: Schrier D J , Sternick J L , Allen E M , Moore V L
Published: December 2022

The genetics of BCG-induced anergy was studied in mice by evaluating delayed hypersensitivity to sheep erythrocytes. Data obtained using congenic mice and by linkage analysis suggested that genes linked to the H-2 complex do not influence the development of anergy. However, studies in allotype-congenic partners (BALB/c; BALB.Igb) indicated that anergy was influenced by genes linked to the immunoglobulin heavy chain complex (Igh). Breeding studies indicated that the gene influencing anergy is recessive. These studies were extended by using BXD recombinant inbred mice, and verified that anergy is controlled by genes linked to the Igh complex. Because several markers and a tentative map has been constructed for the Igh complex on chromosome 12 in BXD mice, we suggest that the gene that controls BCG-induced anergy is located on the centrometric side of Igh-C approximately 18 to 28 recombination units.
Repertoires of T cells directed against a large protein antigen, beta-galactosidase. I. Helper cells have a more restricted specificity repertoire than proliferative cells.
Authors: Krzych U , Fowler A V , Miller A , Sercarz E E
Published: December 2022

The proliferative and helper T cell repertoires were compared in the CBA/J mouse for the response to the large protein antigen, tetrameric beta-galactosidase (GZ = 1021 a.a/monomer). The systems assessed the ability of cyanogen bromide (CB) peptides of GZ to: 1) prime for a T cell proliferative response to GZ; or 2) generate T cell help, measured by the production of anti-FITC PFC in the in vitro response to GZ-FITC. Priming for in vitro proliferation was attempted with 11 CB peptides comprising 70% of the GZ molecule. Strong priming was found with five peptides and intermediate priming was found with four other peptides; two peptides were without effect (CB-20 = a.a 767-862, and CB-4 = a.a. 188-202). Despite this indication of generally dispersed recognition of GZ epitopes, only two CB peptides, CB-2 (a.a. 3-92) and CB-10 (a.a. 378-418) were able to induce a T helper cell response. The surprising dearth of helper T cell-inducing epitopes may be peculiar to the limited fluorescein (FITC) substitution on GZ-FITC (17-25 FITC residues per tetramer) or it may reflect the constraints involved in T cell recognition required for T-B collaboration. Also considered was the possibility that the helper T cell repertoire might be distinct from the proliferative repertoire, the latter reflecting DNA synthesis and recruitment by other functional T cell subpopulations.
Antigenic requirement for induced rejection of long-surviving murine heart transplants.
Authors: Burdick J F , Russell P S
Published: December 2022

Primarily vascularized B10.BR mouse cardiac allografts survive long-term in untreated B6AF1 recipients. We found that sudden complete rejection of the grafts was produced by sequential "triple treatment" with BCG, cyclophosphamide, and a small number of donor-strain lymphocytes. Neither the graft itself, nor donor-strain platelets or frozen-thawed lymphocytes were an effective substitute for the intact donor-strain lymphocytes. Thus the long-surviving graft presents target antigens but lacks the antigenic characteristics required to stimulate a rejection response that are expressed on lymphocytes.
Helper cell factors restore antibody responses of allogeneic bone marrow chimeras: evidence for ineffective cellular interactions.
Authors: Coico R F , Krown S E , Good R A , Hoffmann M K
Published: December 2022

We investigated the nature of deficient antibody responses to SRBC in stable, fully allogeneic bone marrow chimeras. No evidence for a suppressor cell-mediated mechanism was found. Chimera spleens possessed adequate numbers of antigen-reactive B cells to produce a normal antibody response. Using separated chimera cell populations and soluble helper factors, we assessed the functional capabilities of chimera B cells, T cells, and macrophages. Our data suggest that the failure of allogeneic chimeras to produce antibody is not the result of impaired B cell, T cell, or macrophage function, but rather that it is due in ineffective cellular interactions that normally result in the generation of helper factors. In vitro stimulation of chimera macrophages with LPS, and of chimera spleen cells with Con A, resulted in the release of soluble helper factors that were capable of fully restoring chimera B cell responses.
Monoclonal antibody against human interleukin 2 (IL 2). I. Purification of IL 2 for the production of monoclonal antibodies.
Authors: Stadler B M , Berenstein E H , Siraganian R P , Oppenheim J J
Published: December 2022

Human interleukin 2 (IL 2) was produced under serum-free conditions by stimulating mononuclear cells with concanavalin A (Con A) in the presence of phorbol myristate acetate (PMA) and hydroxyurea. The IL 2 was partially purified by sequential chromatography by using phenyl-Sepharose, DEAE Sephacel, and AcA 54 gel filtration. This partially purified material was used to immunize BALB/c mice. After immunization with a total of 48,000 U (spec. act. approximately 10(5) units/mg protein), the spleen cells were adoptively transferred into x-irradiated syngeneic mice and the animals were boosted with another 12,000 U of IL 2. Four days later their spleen cells were hybridized with plasmacytoma cells. Supernatants of the hybridoma cultures were screened for their capacity to inhibit the IL 2-induced proliferation of the CT6 cell line. After expansion and cloning eight different lines were selected for ascitic antibody production. The monoclonal antibodies inhibited the proliferation of the IL 2-dependent cell line in response to either human crude or purified IL 2, as well as rat and mouse IL 2. However, these anti-IL 2 antibodies did not inhibit the proliferation of human T cell lines capable of producing IL 2. Monoclonal antibodies coupled to Sepharose 4B absorbed IL 2 crude culture supernatant, confirming that they react directly with IL 2. The absorbed IL 2 could, for the most part, be eluted by using sodium dodecyl sulfate, thus providing a means for further immunoaffinity purification of IL 2.
Characterization of the human cellular immune response to purified group A streptococcal blastogen A1.
Authors: Regelmann W E , Gray E D , Wannamaker L W
Published: December 2022

The human mononuclear cell response to purified extracellular streptococcal protein, blastogen A, was compared to the response of these cells to PHA and tetanus antigen. Blastogen A induced peak uptake of thymidine during day 6 of tissue culture whereas PHA induced peak uptake during day 5 and tetanus during day 8. Like PHA, blastogen A transformed human umbilical cord lymphocytes and those of nonimmune animals. Also like PHA, blastogen A transformed primarily T lymphocytes. However, unlike PHA, the ability of T lymphocytes to respond to blastogen A was almost completely dependent on the presence of viable non-T lymphocytes. Monocytes were not as effective in facilitating the response to blastogen A as they were for PHA. Thus, blastogen A behaves most like a polyclonal T lymphocyte mitogen, although the degree of dependence of the transformation response on the presence of non-T lymphocytes is much greater than that of PHA.
Communicative interactions between subpopulations of human T lymphocytes required for generation of suppressor effector function in a primary antibody response.
Authors: Morimoto C , Distaso J A , Borel Y , Schlossman S F , Reinherz E L
Published: December 2022

The cellular interactions necessary for generation of human T suppressor effector function were examined in a primary in vitro antigen-specific anti-DNP antibody-forming system. During the induction of the anti-DNP response, it was found that T cells of both T4+ and T8+ subsets were necessary to suppress antibody formation, whereas after activation, only the T8+ subset was required. Thus, a population of T4+ T cells appears to be necessary to activate or induce a subset of T8+ cells to suppress. The T4+ suppressor inducer population, like the resting or activated T8+ suppressor effector subset, was sensitive to low dose irradiation. Moreover, the radiosensitive T4+ subset and the previously defined T4+JRA+ subset were shown to be functionally similar or complementary in that both were required for generation of suppressor effector function. These findings suggest that T-T interactions between radiosensitive T4+JRA+ T cells and radiosensitive T8+ T cells are necessary for suppression of primary antigen-specific antibody production in man.
In vitro alternative pathway activation of complement by the brush border of proximal tubules of normal rat kidney.
Authors: Camussi G , Rotunno M , Segoloni G , Brentjens J R , Andres G A
Published: December 2022

The purpose of this study was to investigate which structures of the nephron, if any, are capable of directly activating the complement (C) system. To this end, two sets of experiments were performed. First, activation of C was assessed on sections of frozen kidney tissue, using the indirect immunofluorescence technique for the demonstration of C fixation. Second, glomerular or tubular fractions of kidney were incubated with normal fresh serum, and subsequent C consumption was measured. The data obtained support the interpretation that the brush border of proximal tubules activates the alternative pathway of the C system. This phenomenon may have pathogenic significance in conditions of aselective proteinuria.
High molecular weight lymphocyte surface proteins are structurally related and are expressed on different cell populations at different times during lymphocyte maturation and differentiation.
Authors: Sarmiento M , Loken M R , Trowbridge I S , Coffman R L , Fitch F W
Published: December 2022

HMW lymphocyte surface proteins T20 and B220, which are expressed on T and B cells, respectively, have been found to be extensively related in structure, beyond what would be expected by the sharing of the determinant detected by monoclonal antibody 13/2. In addition, another HMW protein (T200A) has been discovered, which is preferentially expression proliferating T cells, and which exhibits significant structural homology with T200, but does not express the 13/2 determinant. T200A bears a novel antigenic determinant, detected by monoclonal antibody BS-S28E, which is not shared by either T200 or B220. A fourth HMW protein (170/100) has been found, which is expressed on most lymphocytes, but which is neither structurally nor antigenically related to T200A, T200, or B220. Monoclonal antibodies reactive with 170/100 inhibit T cell-mediated cytolysis, although monoclonal antibodies reactive with T200A, T200, or B220 do not.
T. cruzi: sensitization to macrophage killing by eosinophil peroxidase.
Authors: Nogueira N M , Klebanoff S J , Cohn Z A
Published: December 2022

In this study, we report that trypomastigotes of T. cruzi coated with eosinophil peroxidase (EPO) become sensitized to killing by normal macrophages that are unable to kill uncoated organisms. EPO bound to the surface of the organisms without affecting their extracellular viability. The intracellular killing of EPO-coated trypomastigotes could be inhibited by catalase and azide, suggesting that toxicity was mediated through the small amounts of hydrogen peroxide generated by the phagocytic event in normal macrophages and the peroxidatic activity of EPO. EPO-coated organisms could be killed in a cellfree system by the addition of H2O2 and either iodide, bromide, or chloride. Omission of H2O2 decreased but did not prevent the killing of trypanosomes by the cellfree system and this residual toxicity was abolished by catalase. This suggests that H2O2 generated by trypanosomes contributes to the death of EPO-coated organisms. EPO-coated organisms could also be killed extracellularly when exposed to normal macrophages at high parasite to cell ratios or when a high phagocytic load of another particle was given simultaneously. This effect could be inhibited by both azide and catalase, but not by superoxide dismutase. This suggests that enough H202 is released by phagocytosis of a high number of organisms to generate toxic concentrations of H2O2 outside the confines of the vacuolar system.
Induction of splenic T cell proliferation by lipid A in mice immunized with sheep red blood cells.
Authors: Mita A , Ohta H , Mita T
Published: December 2022

The stimulatory effect of alkali-treated lipid A on the in vivo proliferation of splenic T lymphocytes was investigated by determining 3H-thymidine incorporation into DNA after cell separation by nylon wool adherence. Our experimental model was the primary immune response against SRBC antigen. Mice received a single i.v. injection of alkali-treated lipid A 2 days post-immunization. The splenocytic IgG-plaque response was enhanced but the IgM-plaque response was not. On the 4th day postimmunization, 2H-thymidine uptake by the B cell population was not amplified. However, the splenocytic DNA content was increased in immunized, adjuvant-treated mice, and the radioactivity count in T lymphocytes of lipid A-treated mice was approximately twice that of lipid-untreated animals. We conclude that alkali-treated lipid A enhances T cell proliferation in mice immunized with red blood cells.
Serologic identification of early members of the T cell lineage.
Authors: Berman J W , Basch R S
Published: December 2022

The SC-1 antigen, identified by thymus-absorbed rabbit anti-mouse brain antiserum, is present on multipotential stem cells. Its presence on cells of the T lineage has been examined by immunofluorescence, and we have demonstrated that it is also a marker for thymic development. Although it is present on the majority of fetal and neonatal thymocytes, its expression declines rapidly and, as early as 1 mo after birth, adult levels are reached. In normal animals, these do not change during adult life, even in mice destined to develop a T cell leukemia. SC-1 does reappear transiently on cells in the regenerating thymus of sublethally irradiated mice. Although the antigen is not associated with preleukemic changes in the thymus, it is expressed on some cells of all spontaneous T lymphomas, and it is uniformly present on tissue culture lines of T cell lymphomas.
Lymphocyte transformation induced by autologous cells. XII. Quantitative and qualitative differences between human autologous and allogeneic reactive T lymphocytes.
Authors: Kozak R W , Moody C E , Staiano-Coico L , Weksler M E
Published: December 2022

The T cell populations that respond to autologous or allogeneic non-T cells are distinct, at least to a considerable extent. This conclusion is based on the differences in a) the size of the two populations; on the average one in 260 (individual frequency range, 1:149 to 1:417) T cells respond to allogeneic non-T cells whereas one in 3100 (individual frequency range, 1:2200 to 1:5000) T cells respond to autologous non-T cells; b) the phenotype of T cells activated in the MLR; autoactivated T cells were drawn predominantly from the OKT4-reactive T cell subpopulation, whereas alloactivated T cells activated in the allogeneic MLR were drawn from the OKT4, OKT5, and OKT8-reactive T cell subpopulations; and c) the specificity of activated and nonactivated T cells obtained on Percoll gradients after autologous or allogeneic MLR. Auto- or allodepleted T cells recovered from the 55 to 60% Percoll interface retained full alloreactivity or autoreactivity, respectively. However, auto- or alloactivated T cells recovered from the 40 to 50% Percoll interface showed secondary kinetics to allogeneic and autologous non-T cells. Whether the alloreactivity or autoreactivity found in autoactivated or alloactivated T cells, respectively, represents true cross-reacting T cells or T cells nonspecifically recruited during the MLR cannot at present be decided.
Ocular immune responses. I. Priming of A/J mice in the anterior chamber with azobenzenearsonate-derivatized cells induces second-order-like suppressor T cells.
Authors: Wetzig R P , Foster C S , Greene M I
Published: December 2022

We studied the cellular immune responses to ocular anterior chamber (AC) priming of mice. A/J mice primed subcutaneously with azobenzenearsonate-coupled spleen cells (ABA-SC) manifested delayed-type hypersensitivity (DTH) in the form of footpad swelling when challenged 5 days later with the diazonium salt of ABA. Mice inoculated with ABA-SC in the anterior chamber at the time of subcutaneous priming, however, were tolerant to ABA. Subconjunctival inoculation with ABA-SC did not tolerize; rather it primed for DTH. Antibodies against ABA were not detectable in significant amounts in mice made tolerant by AC inoculation. The AC-induced tolerance was shown to result from hapten-specific T cell-mediated suppression. Suppressor T cells (Ts) arising from AC priming suppressed the efferent limb of the immune response and did not bear detectable cross-reactive idiotype (CRI) surface receptors. In these phenotypic and functional respects, AC-induced Ts differed from first-order Ts (Ts1) that result from i.v. priming. The results are discussed with respect to immune privilege and the anterior chamber of the eye.
Effect of leucogenenol, a thymothyroid hormone, on allograft rejection and hemolysin formation.
Authors: Rice F A , McCurdy J D
Published: December 2022

Recent investigations in this laboratory have established that daily treatment with leucogenenol, an enolic heterocyclic hormone synthesized by the thymus and/or thyroid, induces neonatally thymectomized mice that have accepted a skin allograft to reject this allograft in 7 to 14 days. Also, daily treatment with leucogenenol causes neonatally thymectomized mice to respond to challenge with sheep erythrocytes with the formation of normal titers of hemolysin. These results demonstrate that the hormone leucogenenol induces the development of those cells, absent in neonatally thymectomized mice, that are necessary for a normal immune response.
The relationship between competence for secretion of H2O2 and completion of tumor cytotoxicity by BCG-elicited murine macrophages.
Authors: Cohen M S , Taffet S M , Adams D O
Published: December 2022

BCG-elicited macrophages from C3H/HeJ and A/J mice secrete H2O2 comparably to BCG-elicited macrophages from control C3H/FeJ mice in response to the pharmacologic stimulant phorbol myristate acetate. By contrast, BCG-elicited macrophages from C3H/HeJ and A/J mice could not effect tumor cytotoxicity, whereas macrophages from the control C3H/FeJ mice could. When BCG-activated macrophages from C57BL/6J mice were held in culture overnight, their ability to secrete H2O2 in response to the pharmacologic trigger declined 40 to 60%; the presence of endotoxin in the overnight cultures did not alter competence for secretion of H2O2. By contrast, the cultured macrophages lost all of their cytolytic competence, but this competence could be fully maintained by the presence of endotoxin in the cultures. In three distinct circumstances, competence for secretion of H2O23 thus did not correlate with competence for completion of tumor cytotoxicity. The data imply that competence for release of H2O2 is not sufficient for completion of tumor cytotoxicity.
Fc receptors of mouse cell lines. I. Distinct proteins mediate the IgG subclass-specific Fc binding activities of macrophages.
Authors: Lane B C , Cooper S M
Published: December 2022

To determine the biochemical basis for the two distinct IgG subclass binding specificities on mouse macrophage-like cell lines, Fc-binding proteins were isolated from P388D1 and J774 cells by affinity chromatography using immobilized IgG. Similar IgG subclass-specific Fc gamma-binding proteins were identified on both cell lines. The Fc-binding proteins were characterized by their IgG subclass binding specificity, size, charge, and trypsin sensitivity. Using immobilized IgG1, a protein was isolated with an average m.w. of 65,000 and a pI range of pH 4.7 to 5.8. This protein could also be isolated, although in reduced amounts, with immobilized IgG2b. Two proteins with average m.w. of 70,000 and 60,000 and a pI range of 3.8 to 4.7 were isolated with immobilized IgG2a. The 60-kd protein appeared to be derived from the 70-kd protein. The IgG1/IgG2b Fc-binding proteins were resistant to trypsin, whereas the IgG2a Fc-binding proteins were sensitive to trypsin. In IgG-binding studies with the isolated proteins, the IgG2a Fc-binding proteins continued to exhibit restricted binding specificity for the IgG2a subclass; however, the IgG1/IgG2b Fc-binding proteins showed a broader specificity and would rebind to IgG1, IgG2a, and IgG2b. The IgG-binding properties and trypsin sensitivity of these Fc-binding proteins were similar to the properties of the IgG binding sites on the intact cells. These data indicate that the IgG subclass-specific binding sites on macrophage-like cell lines are due to the presence of distinct protein molecules on the cell surface.
Mechanisms of protective immunity against Schistosoma mansoni infection in mice vaccinated with irradiated cercariae. II. Analysis of immunity in hosts deficient in T lymphocytes, B lymphocytes, or complement.
Authors: Sher A , Hieny S , James S L , Asofsky R
Published: December 2022

Laboratory mice with genetic or induced immunodeficiencies were vaccinated with irradiated cercariae of Schistosoma mansoni and assayed 4 wk later for their resistance to challenge infection. Athymic nude mice failed to develop immunity to challenge or detectable antibody responses to schistosomula, whereas heterozygote (nu/+) controls and nude mice with thymus grafts displayed highly significant levels of resistance and anti-schistosomulum antibody. Similarly, no resistance or antibody production as observed in vaccinated mice that had been made deficient in B lymphocytes by the injection of anti-mu-chain antisera from birth. In contrast, normal levels of vaccine-induced resistance were observed in mice genetically deficient in the fifth component of complement (C5) and in mice decomplemented before challenge by treatment with cobra venom factor. These results indicate that the resistance to challenge infection induced by irradiated cercariae is both thymus and B lymphocyte dependent and therefore is likely to result from specific immune responses directed against schistosome worms. The data also argue against a role for complement in the effector mechanism of vaccine-induced immunity.
Analysis of the two-signal requirement for precursor cytolytic T lymphocyte activation using H-2Kk in liposomes.
Authors: Herrmann S H , Weinberger O , Burakoff S J , Mescher M F
Published: December 2022

Activation of primed pre-cytolytic T lymphocytes (pCTL) requires two signals: recognition of antigen (signal 1) and interaction with a nonspecific helper factor (signal 2). The two signals necessary for generation of a secondary allogeneic CTL response have been analyzed using H-2Kk in liposomes as the stimulating antigen. Use of the liposomes allows the alloantigen to be separated from responder cells after a brief exposure. Thus, the requirements for effective delivery of each signal could be studied independently. A 12-hr exposure of pCTL to alloantigen was sufficient for optimum signal 1 delivery. pCTL recognition of the antigen occurs during this time, and no requirement for adherent cells could be demonstrated. The structure of the antigen-containing liposomes affects the efficiency of pCTL triggering. Factor(s) necessary for signal 2 could be provided by supernatants from mitogen-stimulated lymphocytes. Alternatively, it could be generated with alloantigen, providing that adherent cells were present. Optimum interaction of factor(s) with pCTL, i.e., optimum delivery of signal 2, occurred only if factor(s) was present at 12 to 24 hr after interaction of pCTL with alloantigen. The results suggest that alloantigen recognition triggers pCTL to synthesize and/or express receptors for the factor(s).
Oscillation of cell surface charge of human peripheral blood lymphocytes after stimulation with concanavalin A.
Authors: Uzgiris E E , Lockwood S H , Kaplan J H
Published: December 2022

Lymphocytes isolated from human peripheral blood were incubated with concanavalin A (Con A), and relative changes in cell surface charge were monitored by measuring cell electrophoretic mobility. Kinetic studies showed a time lag of 2.5 hr between exposure to Con A and development of a significant change in cell surface charge. The possible involvement of de novo synthesis of a factor was suggested by the following observations: 1) addition of the protein synthesis inhibitor, puromycin, abrogated the change in electrophoretic mobility; 2) supernatants obtained from cells incubated with Con A caused a mobility change in a separate group of lymphocytes not previously incubated with this mitogen; 3) supernatants obtained from cell suspensions treated with puromycin were inactive. The mobility response showed an oscillation of membrane charge ranging from a 1- to 1.5-hr period, which eventually disappeared after 8 hr at 37 degrees C. Cells at the minimum of the mobility oscillation were refractory to addition of active supernatants. This suggests the involvement at the lymphocyte surface of some cyclic metabolic process, or perhaps formation of receptor-factor complexes followed by endocytosis of these complexes with subsequent regeneration of the receptors on the cell surface.