bluuebunny/crossref_metadata_2025 huggingface dataset:
T lymphocytes that contribute to the immunoregulation of granuloma formation in chronic murine schistosomiasis.
Authors: Colley D G
Published: December 2022

Immunoregulation of Schistosoma mansoni egg-induced granuloma formation in the mouse occurs as chronic infection is being established. Three different modes of manipulation that are known to affect immuno-regulatory T lymphocytes were applied to this model. Adult thymectomy (AdTx) of CBA/J mice, 6 wk after infection, did not affect the size of their granulomas at 8 or 12 wk after infection. However, by 15 wk after infection, sham-treated mice exhibited continued development of modulation of their granulomas, whereas AdTx mice did not. Hemagglutinating antibody titers against soluble egg antigen preparation were marginally elevated in the AdTx group. Adoptive transfer of the suppressor effect of chronic spleen cells was ablated by AdTx. In vivo administration of 1 ml of rabbit anti-mouse thymocyte serum (RAMTS) over a 3-day period to chronically infected mice resulted in a minimal increase in granuloma formation. This treatment was seen, however, to be efficacious in eliminating the suppression normally associated with adoptive transfer of chronic spleen cells. Hydrocortisone acetate (HyC) treatment of chronically infected mice (5.0 to 7.5 mg per mouse over 3 to 5 days) significantly decreased the size of their granulomas. This diminution may be due to the eosinophil-depleting activity of HyC. Adoptive transfer studies demonstrated that the granuloma suppressor mechanism was HyC-sensitive. A continuing supply of newly developed T lymphocytes appears to be required for the establishment and maintenance of a substantial portion of this immunoregulatory process in chronic infection.
A lymphokine resembling transfer factor that stimulates MIF production by nonsensitive lymphocytes.
Authors: Philp J R , McCormack J G , Moore A L , Johnson J E
Published: December 2022

High potency preparations of a new heat-labile, low m.w. (less than 5000) lymphokine (LMWL) were obtained by culturing tuberculin-sensitive guinea pig peritoneal exudate cells (PEC) with PPD in geometric conditions that promote amplified lymphokine production. This LMWL has the ability, in the presence of PPD, to stimulate nonsensitive PEC to produce a heat-stable molecule(s) resembling MIF with a m.w. in the range 50,000 to 100,000. The effects of the LMWL (less than 5000 daltons) and the MIF-like molecule(s) (50,000 to 100,000 daltons) were defined by the indirect macrophage migration assay and a macrophage deoxyglucose uptake assay. It is possible that LMWL represents a form of transfer factor with the ability to recruit unsensitized lymphocytes to produce MIF.
T cell recognition of Moloney sarcoma virus proteins during tumor regression. I. Lack of a requirement for macrophages and the role of blastogenic factors in T cell proliferation.
Authors: Enjuanes L , Lee J C , Ihle J N
Published: December 2022

The characteristics of the T cell blastogenic response of lymphocytes from Moloney leukemia/sarcoma virus inoculated mice against the virion proteins gp71 and p12 were examined. The proliferative responses to both antigens were independent of a requirement for macrophages as determined by macrophage-depletion experiments, the lack of an effect of antisera against Ia on proliferation and by analysis of the number of cell interactions required for proliferation. The kinetics of antigen-lymphocyte interaction were examined and found to require only 4 to 6 hr for half-maximal activation. Proliferation in response to gp71 or p12 was found to be due to the production of blastogenic factor(s), and using additivity experiments, little if any proliferation appears to be associated with antigen-specific lymphocytes. The production of blastogenic factor(s) does not require proliferation and is temporally displaced from antigen-lymphocyte interaction by 8 to 10 hr.
Activation of first component of complement (C1) in guinea pig serum by a polysaccharide is prevented by C1 inhibitor.
Authors: Schultz D R , Arnold P I
Published: December 2022

A polysaccharide (PS) purified from venom of the ant Pseudomyrmex sp. causes the activation of the classical complement (C) pathway in normal serum, but not in guinea pig serum. To investigate why C was not activated in guinea pig serum, we partially purified guinea pig C1 in the presence of the protease inhibitor p-nitrophenyl, p'-guanidinobenzoate (NPGB). This C1 preparation was activated (mu = 0.15, pH 7.5) by the PS in a dose-dependent reaction after NPGB was eliminated by dilution. The PS decreased the action of the C1 inhibitor for C1 in diluted guinea pig serum, and it also inhibited the activity of highly purified guinea pig C1 inhibitor for C1. There was a direct correlation between the concentration of the guinea pig C1 inhibitor and the loss of ability of the PS to activate C1 in mixtures of constant concentrations of purified guinea pig C1 and purified venom PS, and increasing concentrations of purified guinea pig C1 inhibitor. The activity of the human C1 inhibitor, either in diluted serum or highly purified, was not decreased by the PS. These results show that the PS does not activate guinea pig C1 in serum because its action is blocked by the C1 inhibitor.
In vitro augmentation of rat natural killer (NK) cell activity.
Authors: Reynolds C W , Herberman R B
Published: December 2022

In vitro augmentation of rat natural killer (NK) cell activity was produced by 2 types of treatment. Increased activity occurred "spontaneously" when spleen cells were cultured alone at 37 degrees C. This augmentation was dependent on the presence of adherent, phagocytic cells, presumably macrophages, and was independent of LPS of FCS. Normally low levels of NK activity, present in macrophage-depleted cultured cells, could also be boosted in vitro by incubation with Corynebacterium parvum. This augmentation appeared to be independent of both B cells and macrophages and may be due to stimulation of rat NK cells themselves. Both forms of augmentation were associated with the production of interferon, were found in rats of all ages and strains tested, and should provide an excellent in vitro system for detailed studies of activation of rat NK cells.
Limiting dilution assays for the determination of immunocompetent cell frequencies. I. Data analysis.
Authors: Taswell C
Published: December 2022

A statistical method was developed for the analysis of experimental data from limiting dilution assays. Formulas for the estimation of the frequency of immunocompetent cells within a test population were derived by the statistical methods of weighted averaging, likelihood maximization, and X2 minimization. Equations for the latter 2 were solved by Newton's method of iterative approximation. Estimates obtained by these methods were found to be more valid than those obtained by least squares (LS) fitting as judged by the X2 test and as established by Monte Carlo experiments. X2 minimization was chosen as the preferable estimation method with maximum accuracy and precision (minimum bias and variance) for the standard determination of frequencies; likelihood maximization was used only for the confirmation of results. When data from previously published experiments were reanalyzed, both results and conclusions were found to differ significantly from those originally obtained by LS fitting, thus demonstrating the importance of using proper data analysis methods. In conjunction with the use of available calculators or microcomputers, the method presented here provides a simple and rapid procedure for the valid determination of immunocompetent cell frequencies.
The differentiation of cytotoxic T cells in vitro. IV. Interleukin-2 production in primary mixed lymphocyte cultures involves cooperation between Qa-1+ and Qa-1- "helper" T cells.
Authors: Okada M , Stanton T H , Kuppers R C , Henney C S
Published: December 2022

Studies with C57BL/6-TIaa mice have established that both Qa-1+ and Qa-1- helper T cells are required for the optimal production of Interleukin 2(IL-2) activity in primary MLC. This was established both by depletion of Qa-1 bearing cells by treatment of responder cells with anti-Qa-1 serum in the presence of complement and by positive selection (using FACS II analysis) of those cells displaying Qa-1. Furthermore, microfluoremetry revealed that the great majority of C57BL/6-TIaa and of A/J splenic T cells bore Qa-1 alloantigen but that only those with the highest antigen density were susceptible to complement-mediated lysis.
Ia-positive stimulator cells are required in primary, but not in secondary, mixed leukocyte reactions against H-2K and H-2D differences.
Authors: Minami M , Shreffler D C
Published: December 2022

The expression of I region gene products on stimulator cells in primary and secondary MLR directed against differences determined by the K or D regions of the H-2 gene complex was examined in B10-congenic mice. Treatment of the stimulator cells with anti-Ia antisera and complement (C) eliminates stimulation in primary MLR against K or D region gene products, indicating that the stimulator cells must express Ia antigens on their membranes. Further, stimulation in primary MLR against D region antigens is abolished by treatment of the stimulator cells with C and antisera against either I-A or I-E subregion products, but not by treatment of stimulator cells with C and antisera against I-J subregion products. Therefore, these stimulator cells express on their membranes both I-A and I-E subregion gene products, but not I-J subregion gene products. In contrast, in secondary MLR against K or D region gene products, treatment of stimulator cells with anti-Ia antisera and C does not abolish stimulation, indicating that Ia-negative cells can stimulate primed responder cells. These results indicate that the mechanisms of cell-cell recognition necessary in secondary MLR against K or D region gene products are different from those in primary MLR to the same products.
Glycosylation is not required for membrane localization or secretion of IgM in a mouse B cell lymphoma.
Authors: Sibley C H , Wagner R A
Published: December 2022

We have investigated the importance of glycosylation in determining the function of membrane-bound and secreted immunoglobulin M (IgM). Hickman and Kornfeld (1978) previously observed that glycosylation is required for IgM to be secreted by 104E, a mouse plasma cell tumor. In order to determine whether this requirement is a general one for all forms of IgM, we have used WEHI 279.1, a mouse B lymphoma that synthesizes both the membrane and secreted forms of IgM. In the presence of 5 microgram/ml tunicamycin (Tm), glycosylation of both membrane and secreted IgM is at least 90% inhibited, but total protein synthesis is equivalent in control and Tm-treated cells. Despite the absence of carbohydrate, IgM molecules are properly assembled into monomers for membrane localization. Cells whose surfaces have been stripped of membrane IgM by treatment with anti-mu antibody resynthesize the IgM equally well in the presence or absence of Tm. It is more surprising that the assembly of IgM into pentamers and the secretion of these pentamers into the medium are accomplished at the same rate and to about the same levels in control and Tm-treated WEHI 279.1 cells. This is in sharp contrast to the profound inhibition of IgM secretion observed when the plasmacytoma cell 104E is treated with the same concentration of Tm (5 microgram/ml). Although both WEHI 279.1 and 104E cells secrete IgM, the 2 are models for cells at very different points along the B cell differentiation pathway. The difference in the effect of Tm-treatment on IgM secretion may reflect a cellular change that occurs during this differentiation. The unglycosylated IgM is very sensitive to trypsin digestion, whereas the native forms are not. This suggests that the function of glycosylation may be to stabilize the IgM against proteolysis.
Hapten-specific murine colony-forming B cells: in vitro response of colonies to fluoresceinated thymus independent antigens.
Authors: Pillai P S , Scott D W
Published: December 2022

The ability to clone hapten-specific B cells in agar and to subsequently trigger their clonal progeny to antibody synthesis was investigated. Fluorescein (FL) specific B cells were purified on FL-gelatin dishes and cultured in semisolid agar for 6 to 7 days; individual colonies were then picked for restimulation in microculture. FL-specific B cells could be cloned as efficiently as unpurified splenic B cells. The number of colonies formed depended on the presence of sheep erythrocytes (SRBC) or E. coli lipopolysaccharide (LPS) in the cultures. An additive number of colonies were observed with SRBC + LPS compared to that of SRBC or LPS alone. The colonies obtained from SRBC-containing cultures were stimulatable at high frequency by various FL-conjugated antigens to yield anti-FL PFC. However, colonies grown with LPS as the only additive were not stimulatable by any of the antigens tested. On the other hand, addition of M phi or SRBC as additional "mitogens" along with LPS in the agar resulted in progeny colonies that could respond in vitro. Although M phi did not increase the number of colonies, their presence enhanced the size and in some cases the frequency of stimulatable colonies. These data complement earlier observations in suggesting that different B cell subpopulations may grow under different cloning conditions. Moreover, the ability to stimulate the clonal progeny of single B cells to antibody synthesis should permit further definition of triggering and tolerance events at the single-cell level.
Immune response in experimentally induced uremia. II. Suppression of PHA response in uremia is mediated by an adherent, Ia-negative and indomethacin-insensitive suppressor cell.
Authors: Alevy Y G , Slavin R G
Published: December 2022

The characteristics of the adherent suppressor cell in uremic rats were examined. We found that: 1) adherent spleen cells from uremic rats display a more potent suppressor activity than do control adherent cells; 2) the suppression that is mediated by uremic adherent spleen cells cannot be eliminated by pretreatment with indomethacin, whereas the suppression that is mediated by naturally present adherent cells in the control rat is reversed by pretreatment with indomethacin; 3) the uremic adherent suppressor cell does not have Ia antigens that can be detected by the monoclonal antibodies OXL, whereas control adherent cells have Ia antigens on their surface; 4) both the control and uremic adherent suppressor cells are insensitive to mitomycin C and do not have any detectable levels of Thy 1 antigens on their surface. It appears that immune suppression in uremic rats is mediated by an adherent cell that differs from adherent cells present in control animals. The suppression in uremic rats is either not mediated by prostaglandins or may be mediated by preformed prostaglandin synthetase products.
Multiple expression of Ig lambda-chain encoding RNA species in murine plasmacytoma cells.
Authors: Schwartz R C , Sonenshein G E , Bothwell A , Gefter M L
Published: December 2022

The murine plasmacytoma MOPC 315, which synthesizes a complete IgA molecule containing lambda 2 light chains, is shown to synthesize a polypeptide related to the lambda 1 light chain. The abnormal lambda 1 polypeptide (m.w. approximately equal to 18,000) is encoded by a shorter polyA+ mRNA than the lambda 1 mRNAs of lambda 1-producing plasmacytomas. Polypeptides containing lambda 1-specific antigenic determinants have been found to be synthesized both in vitro by translations of RNA and in intact cells. HOPC-1, a lambda 1-producing plasmacytoma, was found to synthesize, in addition to the normal lambda 1-encoding mRNA, a lambda 2 RNA of normal length. A lambda 2-producing hybridoma does not appear to synthesize RNA encoding lambda 1 sequences. The results presented here show that more than 1 light chain gene may be transcribed within a single plasmacytoma cell. Although more than 1 light chain gene may be transcribed, the product of only 1 gene appears to be utilized for the synthesis of complete immunoglobulin molecules. This suggests novel mechanisms for the control of immunoglobulin expression.
In vitro induction of tumoricidal and suppressor macrophages by lymphokines: possible feedback regulation.
Authors: Taramelli D , Holden H T , Varesio L
Published: December 2022

Proteose-peptone-induced macrophages (pM phi) from C57BL/6 (B6) mice were treated in vitro with lymphokine, and 2 functions were evaluated: the ability to suppress lymphokine production and the cytotoxic capacity against tumor target cells. When 10% to 20% pM phi treated with lymphokine were added to a lymphokine-producing system, strong inhibition of MIF and MAF production occurred. The suppression was not specific, since pM phi inhibited the production of lymphokine obtained by either mitogenic stimulation of normal spleen cells (NSC) or alloantigen stimulation of immune spleen cells (ISC). The suppressor cells were adherent, phagocytic, Thy 1.2 negative, with monocyte-macrophage-like morphology. Lymphokine-treated pM phi were also tumoricidal when tested in 18 hr microcytotoxicity assay against RL male 1 lymphoma target cells. However, using endotoxin-free reagents (less than or equal to 0.01 ng/ml of endotoxin by the LAL assay), we found that small amounts of lipopolysaccharide (LPS) were required, in addition to lymphokine, to induce tumoricidal activity in pM phi. In contrast, noncytotoxic pM phi stimulated with lymphokine alone were also able to inhibit lymphokine production, although not as effectively as pM phi stimulated by lymphokine plus LPS. Therefore, we concluded that lymphokine treatment itself is sufficient to induce M phi to become suppressor cells, whereas an additional signal is necessary to induce cytolytic activity. We suggest that there is a negative feedback mechanism of control on the lymphokine-producing cells through the activation of suppressor M phi by lymphokine.
Serum antibody-binding antibodies produced during a primary antibody response.
Authors: Cowdery J S , Steinberg A D
Published: December 2022

By using a solid phase radioimmunoassay we have demonstrated that immunization with either polyinosinic.polycytidylic acid (rl.rC) or TNP-Ficoll (both Tl-2 antigens) leads to the late production (day 10 to 15) of an IgG anti-antibody with specificity for the early (day 4 to 5) IgM produced in response to primary immunization. Adsorption of late sera with antigen removes anti-rl.rC or anti-TNP but does not remove anti-antibody. These results show that specific anti-antibody is found in serum during a primary immune response, and its appearance is coincident with a fall in serum anti-antigen. This suggests a possible role of autologous anti-antibody in regulation of a primary immune response.
Alternative induction of IFN-alpha and IFN-gamma by Listeria monocytogenes in human peripheral blood mononuclear leukocyte cultures.
Authors: Nakane A , Minagawa T
Published: December 2022

Production of interferon (IFN) by Listeria monocytogenes (LM) in human peripheral blood mononuclear leukocyte (PBML) cultures was studied. Acid-labile IFN-gamma was produced in Ficoll-Hypaque-purified mononuclear cell cultures in response to heat-killed LM (HK-LM). However, plastic dish (PD)-nonadherent cells produced acid-stable IFN-alpha, whereas the cells reconstituted with PD-adherent cells (M phi) and PD-nonadherent cells produced IFN-gamma by HK-LM, IFN-gamma was produced only in a mixture of SRBC rosette-forming (T) cells and M phi. Neither T cells nor M phi produced IFN by HK-LM. On the other hand, IFN-alpha was produced from non-T and nylon wool-nonadherent (non-T, non-B) cells by stimulation with HK-LM in the absence of M phi. A reciprocal type of IFN produced in human PBML was absolutely dependent on the presence or absence of M phi. A soluble fraction extracted from sonically disrupted LM had a high potential to induce IFN-gamma in human PBML. Possible mechanism of the alternative production of IFN-gamma or IFN-alpha in human PBML by Listeria are discussed.
Isolation and partial characterization of a 72,000-dalton glycoprotein (Tgp72) on human thymus and T cells: possible relationship to mouse Ly-1 antigens.
Authors: Ishii Y , Fujimoto J , Koshiba H , Kikuchi K
Published: December 2022

A xenoantiserum was raised in guinea pigs to a human T cell antigen (Tgp72) partially purified from cell membrane of T cell leukemia cells from a patient with chronic lymphosarcoma cell leukemia of T cell-type by using sequential chromatographic procedures. The antiserum reacted with all of human thymus and peripheral T cells as well as with a variety of cultured leukemic T cell lines, but not with B cells, granulocytes, and other cultured lymphoid cell lines, including normal B cell, Burkitt, myeloid, and non-T non-B cell lines. Immunoprecipitated material from 125I-labeled thymocyte, T cell, and T cell leukemia cell-surface glycoproteins by this antiserum was analyzed by SDS-polyacrylamide gel electrophoresis, which disclosed a single component with a m.w. of 72K (Tgp72). Further immunochemical data suggest that Tgp72 antigen on human thymus and T cells is an acidic glycoprotein composed of a single polypeptide rather than sulfhydryl-linked subunits, showing a marked electric charge heterogeneity in DEAE anion-exchange resin chromatography. This is further confirmed by the data obtained from 2-dimensional polyacrylamide gel electrophoresis, indicating that Tgp72 is a 72K m.w. glycoprotein with an isoelectric point ranging from 4 to 7. Because of its similarity in tissue distribution and immunochemical characteristics, Tgp72 on human thymus and T cells was assumed to be a human homologue of Ly-1 antigens in murine system.
Lack of age-associated immune dysfunction in mucosal-associated lymph nodes.
Authors: Szewczuk M R , Campbell R J , Jung L K
Published: December 2022

The magnitude of the immune response in old and young mice to trinitrophenylated bovine gamma-globulin was measured in various lymphatic sites at a cellular level using the plaque-forming cell assay. As we have previously shown, the number of splenic IgM, IgG, and IgA anti-TNP PFC progressively declined in aging C57BL/6J male mice. In addition, mice receiving antigen in the 4 footpads and the base of the tail exhibited similar decline in the number of PFC in the draining peripheral lymph nodes with increasing age. In contrast, mesenteric and mediastinal lymph node IgM, IgG, and IgA anti-TNP PFC response to TNP-BGG in complete Freund's adjuvant, i.p. or via gastric intubation, in old mice remained unimpaired compared with the number in younger mice. The data support the view that the mucosal-associated lymphoid system differs from the systemic system with regard to immune competence with age. Furthermore, the findings imply a site preference for a decline in immune function with aging.
The role of complement in the localization of pneumococci in the splanchnic reticuloendothelial system during experimental bacteremia.
Authors: Brown E J , Hosea S W , Frank M M
Published: December 2022

Pneumococci activate the alternative complement (C) pathway in the absence of demonstrable antibody in normal guinea pig serum. They also activate the classical C pathway in the presence of type-specific antibody and perhaps through other mechanisms as well. A quantitative examination was undertaken of the roles of these 2 pathways of C activation in the splanchnic sequestration of 125I-labeled pneumococci, using a-guinea pig model of pneumococcal bacteremia. Normal unimmunized guinea pigs (NIH-GP) localized more than 3 times as many pneumococci to the liver as the spleen during a period when exponential bloodstream clearance was occurring. C4-deficient guinea pigs (C4D-GP) and cobra venom factor-treated guinea pigs (CVF-GP) showed progressively fewer pneumococci cleared by the liver with concomitant increases in the extent of splenic uptake, demonstrating the important role of C in the clearance of bacteria in the unimmunized animal. Immunization of guinea pigs brought about an increase in pneumococcal sequestration by the liver in NIH and C4D-GP but did not affect the localization pattern of CVF-GP. A comparison of reticuloendothelial system (RES) localization patterns with the rate of removal of bacteria from the bloodstream showed a highly significant correlation between increases in splenic sequestration and persistence of bacteremia. Thus, opsonization by C is an important determinant of the RES clearance of pneumococci. Unlike RBC clearance, where C plus IgM leads to hepatic localization, and C plus IgG tends to produce splenic localization, C in the presence or absence of type-specific antibody tends to cause hepatic localization of pneumococci. When C-mediated opsonic activity is less than optimal, the slower clearance of bacteremia that results is accompanied by an increased dependence on splenic sequestration of pneumococci.
Induction of surface Qa2 on lymphoid cells.
Authors: Rosenson R S , Flaherty L , Reinisch C L
Published: December 2022

Three tumor cell lines isolated from individual murine sarcoma virus-infected BALB/cBy (Qa2-) mice were established. BOMS, ORA I, and Thorbly I were analyzed for the expression of Qa2 and Ly surface differentiation antigens. By Cytofluorograf analysis and absorption techniques, all 3 cell lines were found to be Qa2+, Ly1-, Ly2-. This cell surface phenotype suggests that immature lymphoid cells are uniquely susceptible to transformation by virus. Anomalous appearance of Qa2 on transformed lymphoid cells may be analogous to TL antigen induction.
Isolation and characterization of a mitogenic substance for murine and human B lymphocytes from Ulex europeus seeds.
Authors: Yamaguchi N , Yoshimatsu K , Toyoshima S , Osawa T
Published: December 2022

A mitogenic substance, which stimulates murine and human B lymphocytes, has been isolated from the seeds of Ulex europeus. The m.w. of this mitogen was estimated to be approximately 100,000 by sodium dodecyl sulfate disc gel electrophoresis. Under physiologic conditions Ulex mitogen was in the form of a polymer. The isolated mitogen was found to contain 80% carbohydrate, in which glucose, galactose, and rhamnose were the predominant sugars. The carbohydrate portion of the mitogen could be responsible for the mitogenic activity, since the activity was destroyed by periodate treatment but not by protease digestion. It is unlikely that the mitogenic activity is due to the contamination by lipopolysaccharide (LPS), because Ulex mitogen strongly activated spleen cells from C3H/HeJ mice and did not contain a detectable amount of fatty acid. Spleen cells from nude mice, T-depleted spleen cells from conventional mice, or T-depleted spleen cells from conventional mice, or T-depleted lymphocytes from human peripheral blood responded as strongly as lymphocytes (T + B) from normal origin as measured by 3H-thymidine incorporation. It seems that the mitogenic effect is primarily on B cells. Ulex mitogen also induced the increase of immunoglobulin synthesis in murine and human B cells.
Lymphoid cell surface receptor for Moloney leukemia virus envelope glycoprotein gp71. II. Isolation of the receptor.
Authors: Schaffar-Deshayes L , Choppin J , Lévy J P
Published: December 2022

The lymphoid cell surface receptor for Moloney leukemia virus envelope glycoprotein gp71 was isolated by using anti-gp71 antibodies to immunoprecipitate the receptor-gp71 complex from detergent extract of radiolabeled murine thymus cells. Upon sodium dodecyl sulfate gel electrophoresis, a single molecule with an apparent m.w. of 190,000 was identified as the putative gp71-receptor. Analysis in nonreducing conditions indicated that the receptor may be composed of at least 2 subunits of 190,000 daltons.
The use of a monoclonal anti-idiotype antibody to study the biology of a human B cell lymphoma.
Authors: Hatzubai A , Maloney D G , Levy R
Published: December 2022

Immunoglobulin was obtained from the tumor cells of a patient with nodular lymphoma by hybridization to mouse myeloma cells. The human immunoglobulin secreted by these hybridoma cells was used as an immunogen to make murine monoclonal antibodies. Antibodies specific for idiotype, mu heavy chain and lambda light chain, were produced. One anti-idiotype antibody was used to document that idiotype-positive cells and low levels of 19S IgM idiotype were present in the patient's blood. The levels of each were found to correlate with the patient's disease activity. The monoclonal anti-idiotype was effective in eliminating idiotype-positive cells in vitro by solid phase absorption or by complement-mediated cytotoxicity. The anti-idiotype was also used to analyze the host's immunologic response to his own tumor idiotype. There was neither a detectable anti-idiotype antibody response produced in vivo nor a detectable population of T cells that expressed idiotype of could bind idiotype.
A kinetic analysis of immune-mediated clearance of erythrocytes.
Authors: Meryhew N L , Runquist O A
Published: December 2022

A mathematical expression has been derived that successfully correlates the kinetic data for the immune-mediated clearance of red blood cells. The expression resulted from the solution of differential equations arising from a clearance mechanism that was, essentially, consistent with that described by Schreiber and Frank. The mathematical expression correlated data for both IgG-and IgM-mediated reactions. Four different rate constants appear in the final kinetic equation; these constants, which measure the rates of the various steps in the clearance process, were evaluated by an iterative curve-matching process. The values of the rate constants were found to be dependent upon type of sensitizing immunoglobulin, number of C1-fixing sites, and several known immune system modifiers. Correlation of the derived rate expression with the experimental data provided a critical test for the Schreiber-Frank mechanism and the values of the rate constants provided additional insights into the immune clearance process.
The regulation of growth and differentiation of a murine B cell lymphoma. I. Lipopolysaccharide-induced differentiation.
Authors: Boyd A W , Goding J W , Schrader J W
Published: December 2022

The B cell lymphoma WEHI 231 has been previously characterized as resembling a resting B cell, bearing large amounts of IgM on the cell surface, but not secreting immunoglobulin. When WEHI 231 cells were cultured together with low concentrations of lipopolysaccharide (LPS) (0.01 to 3 micrograms/ml), increased levels of immunoglobulin were detected in culture supernatants. Analysis by immunoprecipitation and gel electrophoresis demonstrated that this was secreted (19S) IgM. It was noted that small amounts of secretory IgM were produced even in control cultures not deliberately exposed to LPS. Analysis of the cell lysates showed that LPS treatment resulted in a reduction of synthesis of the surface form of IgM. In contrast, the cytoplasmic pool of IgM precursors was considerably expanded by LPS treatment, reflecting the overall increase in synthesis of IgM in LPS-treated cells. These changes in IgM metabolism appear to parallel closely those occurring in normal B cells after mitogen or antigen challenge. This cloned tumor line promises to be a valuable system in which to investigate some of the molecular events in B cell differentiation.
Characterization of macrophage subpopulations responsive to activation by endotoxin and lymphokines.
Authors: Lee K C , Wong M , McIntyre D
Published: December 2022

The subpopulations of murine macrophages (M phi) that can be activated by endotoxin (LPS) and lymphokines to produce lymphocyte-activating factor, or interleukin 1 (IL-1), and to express cytostatic and cytolytic activities against tumor cells were characterized in 3 cell populations with immature and mature M phi in various proportions, namely, normal peritoneal cells, glycogen-induced peritoneal cells, and cultured bone marrow M phi. There is disagreement in the literature as to whether tumor cytotoxicity can be elicited from all M phi or from a subset of small, newly formed peroxidase-positive (p+) monocytes. Using the technique of cell size fractionation, which separates p+ and p- M phi to various extents, we have found that the magnitude of te response to LPS and lymphokines increases with cell size (and hence maturity or the degree of prior stimulation) and is not exclusive to p+ M phi. Our results are consistent with the concept that all M phi, irrespective of developmental stage, can exist in a continuum of activated states, depending on the nature of the activation signals.
Lymphokine maintains macrophage activation for tumor cell killing by interfering with the negative regulatory effect of prostaglandin E2.
Authors: Taffet S M , Pace J L , Russell S W
Published: December 2022

Mouse resident peritoneal macrophages activated with bacterial lipopolysaccharide (LPS) rapidly lost their ability to kill tumor cells in vitro. Such loss of killing has previously been attributed to the effects of prostaglandin E (PGE) produced by the LPS-stimulated macrophages. Macrophages exposed in the current study to both LPS and partially purified lymphokine did not lose cytolytic activity, in spite of the fact that these cells produced undiminished amounts of PGE, compared to controls. Cytolytic activity was shown to be retained under these conditions because lymphokine decreased the sensitivity of activated macrophages to the negative regulatory effects of PGE. The mechanism responsible for the lymphokine effect is not known; however, generalized inhibition of macrophage responsiveness to the hormone does not appear to be involved because lymphokine did not reduce the cyclic AMP response of macrophages, measured on a whole cell basis, after they were exposed to PGE.
Prevention of experimental allergic encephalomyelitis by bacterial lipopolysaccharides: inhibition of cell-mediated immunity.
Authors: Raziuddin S , Kibler R F , Morrison D C
Published: December 2022

The immunization of Lewis rats with bacterial lipopolysaccharides (LPS) precomplexed to guinea pig myelin basic protein (BP) in complete Freund's adjuvant inhibits the development of experimental allergic encephalomyelitis (EAE) in these animals. These protected animals fail to manifest significant in vivo delayed-type hypersensitivity skin tests and in vitro lymphocyte proliferative responses to BP. Our results indicated that LPS induces a nonspecific reduction in immune reactivity of BP in Lewis rats.
Naturally occurring antibodies to liposomes. II. Specificity and electrophoretic pattern of rabbit antibodies reacting with sphingomyelin-containing liposomes.
Authors: Strejan G H , Essani K , Surlan D
Published: December 2022

Sera obtained from rabbits after immunization with a variety of unrelated antigens contain antibodies that induce complement- (C) mediated lysis of sphingomyelin-containing liposomes in the absence of the relevant antigen from the membrane. Absorption or inhibition with dimyristoyl-phosphatidyl choline-containing liposomes were less effective than with sphingomyelin-containing liposomes in decreasing or abolishing C-dependent lysis of target-liposomes. Phosphoryl choline chloride inhibited the C-dependent lysis mediated by these antibodies, but only when used in high molar excess and in the presence of low antibody concentrations. Purified anti-liposome antibodies displayed an isoelectric focusing pattern consistent with a polyclonal response. The findings confirm the antibody nature of the anti-liposome activity of rabbit sera and indicate that their predominant specificity is directed against conformations of the phospholipid molecule in which the polar (phosphoryl choline) group does not have a major contribution.
Characterization of a monoclonal antibody (5E9) that defines a human cell surface antigen of cell activation.
Authors: Haynes B F , Hemler M , Cotner T , Mann D L , Eisenbarth G S , Strominger J L , Fauci A S
Published: December 2022

This study describes the monoclonal antibody 5E9 and the cell surface antigen it defines. The hybridoma cell line T3-5E9 was derived from fusion of P3 X 63/Ag8 myeloma cells and spleen cells from BALB/c mice immunized with HSB-2 cells, a human T cell line. Although binding to only 1 to 5% of peripheral blood (PB) and spleen mononuclear cells, 5E9 antibody bound to 40 to 80% of Con A-, PHA-, or PWM-activated PB cells. Moreover, 5E9 antibody bound to variable numbers of Sezary, acute myelogenous leukemia, and ALL PB leukemia cells. 5E9 antibody bound to all hematopoietic and nonhematopoietic cell lines tested, to 11 +/- 1% of thymocytes, and 40% of nucleated bone marrow cells. Under reducing conditions, immunoprecipitation studies using 5E9 antibody demonstrated 5E9 antigen to be an 90,000 m.w. glycoprotein. Under nonreducing conditions, antigen 5E9 is a disulfide-linked dimer of approximately 190,000 daltons. Sequential precipitation experiments using antibody 5E9, alpha OD heteroantiserum (raised against T ALL cells), and monoclonal antibody OKT9 demonstrated that the 3 antibody preparations recognized the same 90,000 m.w. glycoprotein. Thus, antibody 5E9 defines an 90,000 m.w. human cell surface antigen that is absent on the majority of PB mononuclear cells and is expressed on rapidly dividing normal and malignant human cells. This monoclonal antibody should be a useful marker of human cell activation.
T lymphocyte responses to non-H-2 histocompatibility antigens. I. Role of Ly-1+2+ T cells as cytotoxic effectors and requirement for Ly-1+2+ T cells for optimal generation of cytotoxic effectors.
Authors: Wettstein P J , Frelinger J A
Published: December 2022

Cytotoxic effector T cells specific for non-H-2 histocompatibility (H) antigens were examined for phenotypic expression of lymphocyte differentiation (Ly) antigens. Virtually all H-Y-specific cytotoxic effectors generated in mixed lymphocyte culture were Ly-1+2+ T cells. H-3-specific effectors comprised both Ly-1+2+ and Ly-1-2+ T cells. However, cytotoxic effectors specific for multiple non-H-2 H antigens were predominantly Ly-1-2+ T cells. The optimal generation of H-Y- and H-3-specific effectors required Ly-1+2+ T cells; optimal generation of multiple non-H-2 H antigen-specific effectors required an interaction between Ly-1+2- and Ly-1-2+ T cells. These observations suggest that the identity of the target H antigen in part determines the Ly type of responsive T cells. Our observations suggest that 2 alternative pathways of T cell response exist for non-H-2 H antigens. The first pathway involves an interaction between Ly-1+2- helper T cells and Ly-1-2+ cytotoxic effector precursors. The 2nd pathway simply involves the response of Ly-1+2+ T cells proliferating and generating H antigen-specific cytotoxic effectors.
Synergy between IgG and monoclonal IgM antibodies in antibody-dependent cell cytotoxicity.
Authors: Shen L , Lydyard P M , Roitt I M , Fanger M W
Published: December 2022

Hybridoma-derived mouse monoclonal IgM anti-ox erythrocyte (ORBC) antibodies, which differed in their ability to exhibit complement-mediated cytotoxicity, direct agglutination and sensitization of indicator cells for detection of receptors for IgM on erythrocytes, failed to mediate antibody-dependent cell cytotoxicity (ADCC) by human K cells. Polyclonal rabbit IgM anti-ORBC antibodies were also incapable of mediating ADCC. However, one of the IgM monoclonal antibodies studied (clone 100) was highly efficient in synergizing with IgG in ADCC, especially when the E rosette-forming enriched fraction was used as a source of effector cells. This synergy was greater after overnight incubation of the lymphocytes and was partially blocked by human IgM. Since all the monoclonal antibodies studied bound efficiently to target ORBC as determined by the additional binding of 125I-labeled anti-k antibodies, the synergy did not appear to be due to a quantitative difference in the binding capacity of the antibodies. That the synergistic effect of purified clone 100 IgM was not due to heterophile IgG antibody was indicated by its sensitivity to reduction and lack of binding to a protein A column. The data are discussed in terms of the possible in vivo relevance of IgG-IgM synergy to immune protection, especially during the development of a primary antibody response when limiting amounts of specific IgG antibodies could be made more effective with the more abundant IgM antibodies.
Partial purification and chemical characterization of macrophage cytotoxicity factor (MCF, MAF) and its separation from migration inhibitory factor (MIF).
Authors: Kniep E M , Domzig W , Lohmann-Matthes M L , Kickhöfen B
Published: December 2022

Macrophage cytotoxicity factor (MCF) was purified in 3 consecutive steps including adsorption chromatography on Matrex Gel Red A, hydrophobic chromatography on phenylalanine-Sepharose, and isoelectric focusing. MCF was characterized as a protein with a m.w. of approximately 30,000 by gel filtration on Sephadex G-100 with 2 isoelectric points at 7.4 and 8.4 in the presence of urea. The unpurified supernatant was fairly stable provided that manipulations favoring adsorption to membrane materials used for dialysis or ultrafiltration were omitted. The partially purified preparation was highly unstable. Trypsin treatment did not affect MCF activity, whereas chymotrypsin destroyed it. Treatment with glycosidases and neuraminidase or cultivation of cells in the presence of 2-deoxy-D-glucose or tunicamycin did not impair the MCF activity. MCF was separated from migration inhibitory factor (MIF) by 2 methods: first, isoelectric focusing in the presence of urea, and second by gel filtration on Ultrogel. MCF could be separated from interferon by chromatography on poly(I)-Sepharose.
Differential effect of interferon on the expression of tumor-associated antigens and histocompatibility antigens on human melanoma cells: relationship to susceptibility to immune lysis mediated by monoclonal antibodies.
Authors: Imai K , Ng A K , Glassy M C , Ferrone S
Published: December 2022

Incubation of cultured human melanoma cells with human leukocyte interferon did not change the expression of melanoma-associated antigens (MAA) recognized by monoclonal antibodies and of Ia-like antigens but significantly increased the expression of HLA-A,B antigens and of beta 2-microglobulin (beta 2-mu). The effect is dependent on the dose of interferon and on the incubation time. Interferon-treated melanoma cells showed an increased susceptibility to lysis mediated by monoclonal antibodies to HLA-A,B antigens and to human beta 2-mu; on the other hand, interferon-treated melanoma cells did not change in their susceptibility to murine natural killer (NK) cell lysis and to immune lysis mediated by monoclonal antibodies to MAA and to Ia-like antigens, and they displayed a reduced susceptibility to human NK cell lysis. Therefore, the increased susceptibility of interferon-treated melanoma cells to lysis mediated by anti HLA-A,B and anti beta 2-mu monoclonal antibodies is likely to reflect the increase in cell surface expression of the corresponding antigens.
Fluid-phase SC5b-8 complex of human complement: generation and isolation from serum.
Authors: Bhakdi S , Roth M
Published: December 2022

A rapid, simple method for isolating SC5b-9 generated in human serum, and SC5b-8 generated in C9-depleted serum is described. The procedure leads to approximately 30% recovery of either complex and consists of polyethyleneglycol precipitation followed by 1 DEAE ion exchange chromatography and a sucrose density gradient ultracentrifugation. The SC5b-8 complex is a water-soluble macromolecular of alpha-electrophoretic mobility. Apart from the absence of C9, it exhibits an identical SDS-gel electrophoresis polypeptide pattern as SC5b-9. A sedimentation coefficient of 19 to 20S and an effective molecular radius of approximately 10 nm were determined by sucrose density gradient centrifugation and gel filtration, respectively. From these values, a m.w. of 800 to 850,000 is tentatively assigned to the SC5b-8 complex. The S-protein was not dissociable from either SC5b-8 or SC5b-9 through treatment with desoxycholate and no alterations in the hydrodynamic properties of either complex were discernible after action of this detergent.
A novel lymphocyte function-associated antigen (LFA-1): cellular distribution, quantitative expression, and structure.
Authors: Kürzinger K , Reynolds T , Germain R N , Davignon D , Martz E , Springer T A
Published: December 2022

We have previously described a monoclonal antibody (MAb), M7/14, which blocks a variety of T cell functions, including CTL-mediated killing, the mixed lymphocyte response, and antigen-specific proliferation. The antigen defined by M7/14 has been designated lymphocyte function-associated antigen one (LFA-1). In this report, LFA-1 has been studied as to cell distribution, surface abundance, structure, and in comparison to other CTL surface antigens, LFA-1 is expressed on lymphoid cells of both the T and the B lineages and on a large fraction of bone marrow cells, but not on exudate macrophages or nonlymphoid tissues. T cells express more LFA-1 than B cells, both in the unstimulated and stimulated states. Compared with unstimulated spleen cells, cytolytic T lymphocyte cell preparations (CTLP) and Con A blasts, but not LPS blasts, show increased LFA-1 expression relative to H-2, and for T cell-containing populations, Lyt-2. M7/14 MAb binds to about 1.5 X 10(4) and 7 X 10(4) LFA-1 sites per average spleen cell or CTLP cell, respectively. M7/14 Mab binds to cTLP in quantitites of 2.5-fold and ies 10.4-fold less than H-2 and Thy-1 Mab, respectively; since the latter have little or no effect on CTL function, inhibition of killing by M7/14 MAb is specific for the LFA-1 surface site. M7/14 MAb and a blocking Lyt-2 MAb are bound in similar quantities of CTLP. LFA-1 is a glycoprotein and consists of 2 noncovalently linked polypeptide chains of 180,000 and 95,000 Mr. The same molecular species as on CTL is present on other T cells and on B cells. The molecular structure and cell distribution of LFA-1 clearly distinguishes it from Lyt-2,3, Ly-5, T145, and T11, which were previously suggested to be either associated with the function of and/or present on the surface of CTL.
Linkage of total deficiency of the second component (C2) of the complement system and of genetic C2-polymorphism to the major histocompatibility complex of the guinea pig.
Authors: Bitter-Suermann D , Hoffmann T , Burger R , Hadding U
Published: December 2022

In the guinea pig six common phenotypes for hemolytically active C2 were detected. They resulted from three allotypic variants: a basic C2B and two more acidic variants C2A and C2A1. (The gene frequency for C2B in the outbred population was 0.36, 0.33 for C2A, and 0.31 for C2A1.) These variants were inherited as autosomal codominant traits. A strong linkage-disequilibrium between C2 and C4 allotypes was observed consisting of the pairs C2A1-C4F. C2A-C4S, whereas C2B was associated with either C4S or C4S1. To investigate the linkage of C2 to the MHC a family of strain 2 X strain OM3 was studied: the C2B allotype segregated with the strain haplotype of OM3 whereas C2A1 does the same with the strain 2 haplotype. During the C2-typing, an animal totally deficient in hemolytic C2 activity was discovered. Its C1, C4, C3, C5, factors B, D, and H were normal. Breeding yielded finally homozygous and heterozygous-deficient individuals. The latter ones exhibited 50 to 60% of the hemolytic activity of normal guinea pigs. Combined typing data of C4, C2 and GPLA showed that the deficient gene behaved like a rare, silent allele of C2, which is GPLA-linked and inherited like the normal C2-variants. An antiserum against C2 was raised in C2-deficient animals, which reacted with normal guinea pig serum but not with serum from C2-deficient animals. No cross-reactivity with human or mouse serum could be detected.