bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytokinetic basis for the impaired activation of lymphocytes from patients with primary intracranial tumors.
Authors: Elliott L H , Brooks W H , Roszman T L
Published: December 2022

Patients with malignant brain tumors have a variety of immunologic abnormalities, including the impaired responsiveness of peripheral blood lymphocytes (PBL) to mitogens and alloantigens. We further investigated this impairment of lymphocyte reactivity by employing the techniques of limiting dilution analysis and cytokinetic analysis. PBL preparations from patients have approximately six times fewer phytohemagglutinin (PHA)-responsive cells than PBL from normal subjects. Similar results were obtained with purified T cell preparations. Cytokinetic analysis of PHA-induced [3H]thymidine incorporation employing colchicine blocking of mitosis demonstrated that the number of first generation cells entering the S-phase of mitosis for each 24-hr period was less for PBL from patients than for PBL from normal individuals. First generation responding cells from patients and normal subjects entered DNA synthesis at the same time (48 to 72 hr). Cytokinetic analysis over a period of 168 hr demonstrated that whereas PBL from normal individuals demonstrated second generation responding cells, PBL from the majority of patients did not, thus indicating a defect in their ability to undergo clonal expansion. Measurement of interleukin 2 (IL 2) activity in culture fluids from PHA-activated PBL from normal subjects and patients revealed significantly lower IL 2 levels in culture fluids from PBL from patients. The addition of various concentrations of lectin-free IL 2 to PBL from patients stimulated with PHA did not restore responsiveness to normal values. There was no difference between the levels of interleukin 1 (IL 1) produced by lipopolysaccharide-activated monocytes from normal subjects and patients. Overall, these results suggest that an intrinsic defect exists in T cells obtained from brain tumor patients that renders them unable to enter into normal mitogen-induced blastogenesis.
Antibodies from patients with mixed connective tissue disease react with heterogeneous nuclear ribonucleoprotein or ribonucleic acid (hnRNP/RNA) of the nuclear matrix.
Authors: Fritzler M J , Ali R , Tan E M
Published: December 2022

The sera of patients with mixed connective tissue disease (MCTD) have high titers of antibodies directed against nuclear U1-ribonucleoprotein (U1-RNP). This antigen is easily extracted from nuclear preparations with physiologic saline and from tissue sections with 0.1 HCl, leaving the nucleic acids and nuclear matrix behind. When U1-RNP is extracted from HEp-2 cells with 0.1 N HCl, the sera of 32/32 patients with MCTD react with another antigen that is exposed by the extraction procedure. This antigen is not destroyed by trypsin and deoxyribonuclease 1 treatment but is sensitive to both purified ribonuclease A and purified micrococcal nuclease. Absorption studies showed that the antibody reacting with this antigen cannot be absorbed by sheep red blood cells coated with extracts of rabbit thymus that contain U1-RNP. Radioimmunoassay showed that the reaction of the unadsorbed antibody was with heterogeneous nuclear ribonucleoprotein or ribonucleic acid (hnRNP/RNA) and not with transfer RNA or ribosomal RNA. The hnRNP/RNA antigen is demonstrated as discrete particles in the internucleolar chromatin of interphase cells, but in metaphase cells the antigen is diffusely dispersed. The distribution, solubility, and biochemical characteristics suggest that the antigenic moiety is part of the nuclear matrix. Therefore, MCTD sera contain antibodies that react with at least two species of nuclear RNP: small nuclear RNP (snRNP), as described by others, and a high m.w. hnRNP/RNA bound to the nuclear matrix.
Prevention of experimental allergic encephalomyelitis (EAE) in the SJL/J mouse by whole body ultraviolet irradiation.
Authors: Hauser S L , Weiner H L , Che M , Shapiro M E , Gilles F , Letvin N L
Published: December 2022

The cellular requirements for the in vivo induction of experimental allergic encephalomyelitis (EAE) were investigated in the SJL/J mouse. Exposure of mice to whole body ultraviolet (UV) irradiation, a treatment that has been shown in other systems to interfere selectively with antigen-presenting cell function, prevented the development of clinical and pathologic signs of acute EAE. Splenic T cells from UV-treated animals did not adoptively transfer resistance to EAE, making it unlikely that UV irradiation resulted in the generation of a specific suppressor cell population responsible for protection from EAE. UV irradiation was effective in preventing EAE when administered before initial immunization; UV irradiation was ineffective in modifying ongoing EAE or in preventing relapses of EAE induced by reimmunization. In additional experiments, adult thymectomized, lethally x-irradiated mice reconstituted with syngeneic marrow cells depleted of mature T lymphocytes were found to be resistant to the induction of EAE. Susceptibility was restored by the addition of splenic T cells, demonstrating that EAE induction is T cell-dependent in the mouse. The prevention of an experimental autoimmune demyelinating disease by whole body UV irradiation suggests that interference with the function of Ia-bearing accessory cells may represent an approach for immunotherapy in autoimmune disorders.
Inhibition of antibody responses to phosphocholine by C-reactive protein.
Authors: Nakayama S , Du Clos T W , Gewurz H , Mold C
Published: December 2022

C-reactive protein (CRP) is an acute phase serum protein in man that binds to the cell wall C-polysaccharide (PnC) of Streptococcus pneumoniae via phosphocholine (PC) determinants. We have previously shown that in mice CRP increases splenic clearance of PnC-coated autologous erythrocytes and S. pneumoniae, and increases survival after pneumococcal infection. Because CRP alters clearance of particulate PnC antigens, we tested its effect on immunization with pneumococci. Pretreatment of mice with 50 to 200 micrograms CRP 30 min before immunization with serotype 3 S. pneumoniae resulted in dose-dependent inhibition of the antibody response to PC. Both serum hemagglutinin and splenic PFC against PC were decreased in CRP-treated mice tested from 1 to 10 days after injection of antigen. CRP treatment had no effect on the antibody response to the serotype 3 capsular polysaccharide, another T-independent antigen. To determine whether CRP inhibition was related to altered processing of particulate antigen, mice were immunized with horse red blood cells (HRBC) conjugated with PC or PnC and the PFC responses to PC and HRBC were determined. CRP treatment resulted in specific inhibition of the PFC response to PC in both cases without affecting the response to HRBC. These results indicate that inhibition of the antibody response by CRP is not the result of altered antigen localization and processing, and that CRP may prevent immunization by masking determinants on bacterial or other surfaces.
Cross-reactive idiotypes on heterologous anti-allotype antibody.
Authors: Roux K H , Metzger D W
Published: December 2022

Specific anti-a1 Ab was isolated from rabbits, guinea pigs, mice, chickens, and a goat. Each of these preparations was able to inhibit the reaction between a1 IgG and rabbit anti-a1 Ab as well as the reaction between rabbit IdX (anti-a1 Ab) and rabbit anti-IdX (anti-anti-a1 Ab). Maximal levels of inhibition ranged from 75 to 100% in the latter assay. Although the relative binding efficiencies of each preparation varied widely, there was generally a positive correlation between the ability of an anti-allotype reagent to bind to a1 IgG and to anti-IdX Ab. Each of the heterologous anti-a1 Ab samples was able to form precipitin bands with rabbit anti-Id Ab. These bands fused with each other and with rabbit anti-a1 Ab. These results weaken the interpretation that the ubiquitous expression of IdX previously observed in the rabbit reflects shared conserved genes. We suggest that either 1) the anti-Id Ab represents high fidelity internal images of the a1 epitopes, or 2) latent a1 allotype Ig is present in the anti-IdX preparation. In either case, any anti-a1 Ab would be potentially reactive with the anti-IdX preparation.
Shared idiotope on monoclonal anti-Ia.7 antibodies reactive with determinants in a structural domain of the I-E molecules.
Authors: Devaux C , Labit C , Marchetto S , Pierres M
Published: December 2022

In previous studies, heterologous anti-idiotypic (anti-Id) antisera against the C3H.SW 14-4-4S or the A.TH 41.A anti-Ia.7 monoclonal antibodies (mAb) were shown to identify an interstrain cross-reactive idiotypic specificity (IdX.Ia.7) expressed on monoclonal or conventional anti-Ia.7 alloantibodies. The objective of the present investigation was to characterize further this IdX at the idiotopic level. To this end, 11 hybridomas producing IgG1, IgG2a, or IgM anti-Id mAb were derived from a rat immunized with a mixture of 10 A.TH or A.BY anti-Ia.7 mAb. The specificity of the latter anti-Id mAb was determined by direct Id binding radioimmunoassay (RIA) with the use of a panel of 52 anti-Ia mAb derived from hybridomas produced in various inbred mouse strains. These rat anti-Id mAb recognized idiotopes expressed on i) all anti-Ia.7 mAb against determinants in the topographic domain I of the I-Ek molecule but not on 18 other anti-I-Ek mAb directed at epitopes in domains II or III; ii) three of 19 anti-I-Ak mAb; and iii) one A.TL-derived anti-I-As mAb. Competitive Id binding assays revealed that among the 14 IdX+ anti-Ia.7 mAb, one (81.B) was bound to a lesser extent by various rat anti-Id mAb, suggesting that heterogeneity probably exists in this antibody family. By contrast, two isologous (B10.S(7R)) anti-Id mAb to the IdX.Ia.7+ mAb 41.A displayed a specificity restricted to 41.A individual idiotopes (IdI). Rat anti-IdX.Ia.7 and mouse anti-41.A IdI mAb inhibited the binding of 125I-labeled mAb 41.A to CBA spleen cells. These two sets of mAb bound in a noncompetitive fashion to mAb 41.A-coated plates, indicating that their corresponding public or private idiotopes were spatially distinct. These data may have implications for in vivo manipulations of anti-Ia immune responses.
Comparison of structural and functional variation in class I HLA molecules: the role of charged amino acid substitutions.
Authors: Monos D S , Tekolf W A , Shaw S , Cooper H L
Published: December 2022

Functional and structural heterogeneity of HLA class I molecules was sought among five donors serologically identical for A2, A3, B7, and Bw44. Functional differences were identified by cell-mediated lympholysis (CML) after allogeneic mixed lymphocyte reaction among the five donors. Structural differences were characterized by high resolution two-dimensional electrophoretic maps of the class I HLA proteins synthesized by peripheral blood lymphocytes of these donors. Cells from three donors showed no CML-defined differences from one another; their HLA protein maps were identical. The cells of one donor recognized an A3-associated target antigen on the cells of all the other donors; her HLA map revealed a unique protein with altered isoelectric point. Another donor's cells differed by two CML-detected antigens: one was identified as a variant of Bw44 ("44.2") and the other was associated with Cw4. This donor's two-dimensional HLA map showed two novel charged proteins. By using these data, a two-dimensional map locating HLA-A2, -A3, -A3', -B7, -Bw44.1, -Bw44.2, and -Cw4 was prepared. Because each of three CML-detected antigens was correlated with a protein of distinctive charge, our results and the available published data raise the possibility that amino acid substitution producing charge variation may be a particularly important mechanism in the generation of CML-detectable HLA diversity.
Residual hemolytic and proteolytic activity expressed by Bb after decay-dissociation of C3b,Bb.
Authors: Fishelson Z , Müller-Eberhard H J
Published: December 2022

Bb (Mr = 63,000) is the catalytic site-bearing subunit of the C3 convertase of the alternative complement pathway, C3b,Bb, which is dissociated from the complex upon decay of the enzyme. Because purified Bb induced certain leukocyte activities, we examined whether it expresses residual hemolytic or proteolytic activity. Hemolytic activity of Bb was tested by using Factor B- or Factor D-depleted normal human serum and rabbit or sheep erythrocytes. Proteolytic activity of Bb was assessed by using purified C3 or C5 as substrates and SDS-PAGE to detect protein cleavage. Bb expressed metal-dependent hemolytic activity that was approximately 100-fold lower than that of Factor B. This activity could be inhibited by Factor H and enhanced by properdin. Low but statistically significant binding of 125I-labeled Bb to C3b on erythrocytes was demonstrated. Monoclonal antibodies that bind to Bb but not to intact Factor B inhibited the Bb hemolytic activity. Purified Bb cleaved C3 to C3a and C3b, as evidenced by the appearance of the alpha'-chain of C3b. It also cleaved C5 to C5a and C5b when cobra venom factor was present in the reaction mixture. Metal ions were required for expression of proteolytic activity, and Ni supported the activity better than Mg. These results indicate that decayed Bb has residual C3 and C5 cleaving activity and hemolytic activity, expression of which appears to require its association with C3b, C3(H2O), or cobra venom factor. These observations may aid in explaining the mechanism of action of Bb on leukocytes.
Characterization of the initial C3 convertase of the alternative pathway of human complement.
Authors: Fishelson Z , Pangburn M K , Müller-Eberhard H J
Published: December 2022

C3(H2O),Bb, the initial C3 convertase of the alternative complement pathway, was demonstrated to be a metal-containing protein complex by sucrose density gradient ultracentrifugation. Demonstration of this labile enzyme became possible by increasing its half-life with nickel instead of magnesium ions used for enzyme formation. The enzyme was generated from C3, the internal thioester bond of which was hydrolyzed (C3(H2O)), and from 125I-Factor B and Factor D. The sedimentation coefficient of the enzyme complex was 10.7S. By using 63Ni for enzyme formation, the metal ion was detected in the enzyme complex after ultracentrifugation in the presence of 10 mM EDTA. The stoichiometry of the constituents in the C3(H2O),Bb(Ni) complex was 1:1:1. To verify that C3 is incorporated into the enzyme complex in the form of C3(H2O), the enzyme complex was adsorbed to anti-Factor B-Sepharose and subjected to decay-dissociation. Examination of the subsequently eluted protein by SDS gel electrophoresis under reducing conditions demonstrated the presence of an intact C3 alpha-chain. This work provides further evidence that a C3 convertase can be generated from noncleaved C3 that is modified at the thioester site. With the use of a fluorometric assay, the activity (kcat/Km) and the half-life of the initial C3 convertase were determined and compared to those of C3b,Bb.
Quinacrine inhibits the primary but not secondary proliferative response of human cytotoxic T cells to allogeneic non-T cell antigens.
Authors: Namiuchi S , Kumagai S , Imura H , Suginoshita T , Hattori T , Hirata F
Published: December 2022

Quinacrine inhibited the generation of cytotoxic T lymphocytes from human peripheral blood T cells stimulated by allogeneic non-T cells as measured by [3H]thymidine incorporation as well as by cytolytic reactions against 51Cr-labeled allogeneic lymphocytes. These observations were further supported by the finding that quinacrine inhibited the expression of Tac antigen, the receptor for growth factor(s). Because other phospholipase A2 inhibitors such as tetracaine and p-bromophenacyl bromide could inhibit these reactions, the inhibition of cytotoxic T lymphocyte responses by quinacrine appeared to be attributable to the inhibition of phospholipase A2 in these cells. In keeping with this interpretation, arachidonate release from phospholipids in cytotoxic T cells stimulated by allogeneic non-T cells was inhibited by quinacrine. In contrast, the pretreatment of T cells with quinacrine did not result in the inhibition of their secondary proliferative response to the same stimulation with allogeneic non-T cells. These results, taken together, suggest that quinacrine does not inhibit the recognition of antigens by cytotoxic T cells, while it blocks the mitogenic response of T cells to allogeneic antigens.
Schistosome egg antigen(s) presentation and regulatory activity by macrophages isolated from vigorous or immunomodulated liver granulomas of Schistosoma mansoni-infected mice.
Authors: Elliott D E , Boros D L
Published: December 2022

Previously, it was shown that macrophages isolated from liver granulomas of Schistosoma mansoni-infected mice constituted about 30% of the granuloma cell population, were highly activated, and more than 50% displayed H-2 I region-associated antigens. Moreover, the degree of macrophage maturation/activation correlated with the intensity of the granulomatous response. Thus, inflammatory macrophages from the vigorous granulomas of acutely infected mice (VigGrM phi) displayed greater phagocytic and tumoricidal activity than did M phi from the T suppressor cell-modulated granulomas (ModGrM phi) of chronically infected animals. No difference was seen, however, in the percentage of Ia-bearing M phi isolated from vigorous or modulated liver granulomas. Presently, accessory activity of GrM phi was assayed by reconstitution of the proliferative responses of M phi-depleted lymphocytes. Both VigGrM phi and ModGrM phi were able to reconstitute the Con-A induced proliferation of normal thymoctyes. Moreover, both types of GrM phi could reconstitute parasite egg antigen-specific proliferation of the mesenteric lymph node cells from acutely infected mice. Reconstitution was dependent on M phi that displayed I-A and I-E subregion-encoded determinants. In higher doses the activated VigGrM phi were more suppressive toward lymphocytic proliferation than were the less activated ModGrM phi. Apparently, the degree of M phi suppressor activity was in direct relationship to the state of M phi maturation/activation. These data indicate that the inflammatory M phi of the schistosome granuloma may be involved in the focal immune accessory/regulatory events that occur within these T cell-mediated lesions.
Modified-self-induced modulation of the immune response to herpes simplex virus: effect on antibody formation, cytotoxic T lymphocyte induction, and survival.
Authors: Wilson L A , Karabin J M , Smith J W , Dawson D , Scott D W
Published: December 2022

C3H/HeJ mice were injected i.v. with soluble herpes simplex virus (HSV) envelope antigens coupled to syngeneic splenocytes 7 days before challenge i.p. with either soluble HSV antigens in complete Freund's adjuvant or infectious HSV. Fourteen days post challenge, and anti-HSV antibody hyporesponsiveness was observed in the HSV modified-self pretreated animals that was 50% or less than the control response. Both forms of HSV challenge were equally effective. This unresponsiveness was specific for HSV when compared with a concomitant response to chicken red blood cell (CRBC) challenge. Challenge with infectious HSV afforded us the opportunity to investigate the effect of HSV modified-self pretreatment on the induction of HSV-specific cytotoxic T lymphocytes (CTL). In vitro restimulation of in vivo-primed splenocytes with infectious HSV generated HSV-specific CTL. The degree of HSV-specific CTL response was found to be variable in animals that received the HSV modified-self pretreatment with respect to control animals. Survival was significantly greater in HSV modified-self pretreated mice after infectious HSV challenge than in control animals.
Manipulation of anti-LDH-B response by T suppressor factors.
Authors: Ikezawa Z , Nagy Z A , Klein J
Published: December 2022

Hybridomas produced by fusion between the BW5147 thymoma and an LDH-B-specific B10.A(2R) suppressor T cell line secrete two T suppressor factors (TsF). One factor (TsF-A) shares Mhc determinants with the A alpha A beta molecule and suppresses proliferating Th cells; the other (TsF-E) shares determinants with the E alpha E beta molecule and it inhibits the maturation of the T suppressor (Ts) cells. Here we demonstrate that the two factors can be used to alter the immune response status of cultured T lymphocytes or of an animal. When added to a culture of LDH-B-primed cells or injected into mice, the TsF-A turns responders into nonresponders, presumably by blocking the proliferation of the Th cells. The TsF-E converts nonresponder cultures or mice into responders, presumably by preventing the differentiation of Ts cells. As there are good prospects for obtaining TsF in large quantities and in a highly purified form, this manipulation of the immune response by the deployment of specific factors promises to become an efficient new method of immunotherapy.
T cell regulation of immunoglobulin class expression in the B cell response to TNP-Ficoll: characterization of the T cell responsible for preferential enhancement of the IgG2a response.
Authors: Mongini P K , Longo D L , Paul W E
Published: December 2022

Syngeneic T cells injected into athymic nu/nu mice cause a preferential enhancement in the amount of IgG2a anti-TNP Ab produced by these mice to TNP-Ficoll. This enhancement appears to be caused by T cell effects on the IgG switching pathway. Through the use of F1----parent chimeras, the helper T cells were shown to affect TNP-Ficoll-responsive B cells in an H-2-unrestricted manner. The ability of T cells to mediate this IgG2a enhancement did not appear to be unique to any particular murine genetic background, because it was observed with T cells and nu/nu mice of C57BL/10, BALB/c, CBA/Ca, and B10.D2 strains. Priming of T cell donors with Ficoll or TNP-Ficoll did not increase the ability of splenic T cells, on a per cell basis, to enhance the IgG2a Ab response to TNP-Ficoll. The T cell population responsible for modulating the isotypic response was found to be sensitive to C-mediated cytotoxicity with both anti-Lyt-2 and anti-Lyt-1 hybridoma Ab. Although T cells from both the thymus and the spleen expressed enhancing activity, splenic T cells were more effective, on a per cell basis, than were thymocytes. The observations suggest that T cells that appear to enhance the switch to IgG2a in TNP-Ficoll-responsive B cells are not effectively primed by the antigen and interact with TNP-Ficoll-activated B cells through an H-2-unrestricted mechanism.
Polyclonal activation of xid B cells by auto-Ia-reactive T cell clones.
Authors: Kimoto M , Yoshikubo T , Kishimoto S , Yamamura Y , Kishimoto T
Published: December 2022

The mechanism of T cell-dependent activation of xid B cells into Ig-producing cells was studied by employing H-2-restricted, antigen-specific T cell clones. Helper factors (B cell stimulatory factors, BSF) released from KLH-specific T cell lines could induce polyclonal Ig production in B cells from (CBA/N X BALB/c)F1 (NBF1) female mice but not from CBA/N or NBF1 male mice. Direct addition of helper T cell lines induced Ig production in xid B cells from CBA/N or NBF1 male mice. A T cell clone, MK6, which was derived from NBF1 male mice and specific against Iad determinant, could activate NBF1 male but not CBA/N B cells. Another clone, CK4, derived from CBA/N mice and having specificity against KLH plus I-Ak determinant could activate both CBA/N and NBF1 male B cells into IgM- and IgG-producing cells in the absence of KLH, and monoclonal anti-I-Ak antibody specifically blocked such activation. These results suggest that xid B cells are able to be activated by the signal provided by the recognition of Ia molecules on B cells by auto-Ia-reactive T cells. Xid B cells from CBA/N mice that had been co-cultured with a T cell line specific against I-Ak determinant for 24 hr became reactive to BSF and capable of differentiating into Ig-producing cells in the presence of BSF. The results showed that even xid B cells could be responsive to BSF if they were in a certain activation stage.
A reappraisal of "T-independent" antigens. I. Effect of lymphokines on the response of single adult hapten-specific B lymphocytes.
Authors: Pike B L , Nossal G J
Published: December 2022

Four supposedly T-independent antigens, fluorescein (FLU)-E. coli lipopolysaccharide (LPS), FLU-Brucella abortus (FLU-BA), FLU-Ficoll, and FLU-polymerized flagellin (FLU-POL), were tested for their capacity to stimulate B cell proliferation and antibody formation. Single, isolated FLU-specific adult murine splenic B lymphocytes were used as the unequivocal target cell in 10-microliter cultures unsupported by accessory, feeder or filler cells. The stimulatory capacity of four supposed mitogens, LPS, dextran sulfate (DXS), BA, and Ficoll, and of one T-dependent antigen, FLU-keyhole limpet hemocyanin (FLU-KLH), was also studied. Stimuli were used over a wide range of concentrations with or without added T cell-derived, antigen-nonspecific factors promoting B cell growth and differentiation (BGDA). The results allowed triggering stimuli to be divided into four groups: 1) FLU-KLH failed to stimulate single B cells even in the presence of BGDA as did unconjugated Ficoll; 2) FLU-POL was stimulatory only when BGDA was also present, as was unconjugated BA; 3) FLU-BA was slightly stimulatory in the absence of BGDA, but much more so in the presence of BGDA, as was a low concentration of LPS; 4) FLU-LPS and FLU-Ficoll were powerfully stimulatory over a 100,000-fold range of concentrations, and at no concentration did BGDA affect their capacity. LPS or LPS + DXS at high concentrations behaved similarly. The differing behavior of the various conjugates did not correlate either with their supposed "TI-1" or "TI-2" status or with the mitogenic properties of the carrier portion of the antigen. For example, FLU-Ficoll at 0.01 microgram/ml caused 28% of FLU-specific B cells to form proliferating clones, 70% of which secreted immunoglobulin, but a 1000-fold higher concentration of Ficoll failed to stimulate single cells. It is tempting to regard stimuli in group 4 as truly T-independent. However, the intentional addition of thymus filler cells to single B cells triggered by these agents markedly raised antibody formation. Until the nature of this further "helper" effect is understood, it would be unwise to ignore the possibility that T cells or accessory cells are involved.
Primary in vitro generation of cytotoxic cells specific for human minor histocompatibility antigens between HLA-identical siblings.
Authors: Tekolf W A , Shaw S
Published: December 2022

A limiting dilution culture system was developed for the primary in vitro detection of human minor histocompatibility antigens by cytotoxic T lymphocytes (CTL). CTL were generated in primary in vitro culture between two HLA-identical sibling pairs and propagated as stable CTL lines. Population and family studies indicate that these CTL lines recognize minor histocompatibility antigens in an HLA-restricted manner. The antigen recognized by one CTL line is detected on six (out of 37) HLA-B7-positive donors but not on 32 HLA-B7-negative donors. The cytotoxicity of this CTL line is mediated by T3+, T8+ effector cells. The antigen detected by this CTL population is different from all known human minor histocompatibility antigens. The data of this study, like those in the mouse system, suggest that a suppressor cell is diluted out in a limiting dilution culture, which allows the activation of the CTL precursors.
Antibodies to glycosphingolipids in patients with multiple sclerosis and SLE.
Authors: Endo T , Scott D D , Stewart S S , Kundu S K , Marcus D M
Published: December 2022

We used a liposome lysis assay to measure antibodies against a panel of glycolipids. Antibodies to one or more compounds were detected in 34 of 46 patients with multiple sclerosis, 19 of 31 patients with systemic lupus erythematosus (SLE), and in the majority of patients with cranial trauma or cerebrovascular accidents. Antibodies against ganglioside GM1 and asialo GM1 were found most commonly, and they were frequently present in the same sera. The specificity of the antibodies was tested in four sera that contained antibodies to both glycolipids. The anti-GM1 antibodies cross-reacted with asialo GM1, but the converse was not true. Among patients whose sera contained antibodies to glycolipids, anti-asialo GM1 alone was more common in patients with SLE (7 of 17) than in multiple sclerosis (2 of 34; p = 0.004). Anti-GM1 alone was found in 9 of 34 patients with multiple sclerosis and 1 of 17 patients with SLE, a difference that was not statistically significant (0.135). No correlation was observed between the presence of anti-glycolipid antibodies and symptoms related to the nervous system in patients with SLE. Because of our inability to detect these antibodies by a solid phase immunoassay (ELISA), a comparison was made of the titers obtained with three monoclonal anti-glycolipid antibodies in the liposome lysis assay and ELISA. The ELISA was less sensitive in all instances, requiring from four to 1000 times as much antibody as the liposome lysis assay to give a positive test. We conclude that antibodies to glycolipids occur frequently in patients with multiple sclerosis, SLE, major cranial trauma, and cerebrovascular accidents. Their role in the initiation or perpetuation of inflammatory disease of the central nervous system has yet to be determined.
Production of human-human hybridomas secreting antibody to sheep erythrocytes after in vitro immunization.
Authors: Strike L E , Devens B H , Lundak R L
Published: December 2022

This report describes the formation of human hybridomas after in vitro immunization of tonsillar lymphocytes and fusion of the stimulated lymphocytes with a human lymphoblastoid cell line, WI-L279HF2. Lymphocytes were stimulated in vitro with sheep erythrocytes (SRBC) and polyclonal activators (PCA). By varying the duration of in vitro immunization before fusion and varying the PCA concentrations, the optimal conditions for cell fusion, hybrid cell growth, nonspecific immunoglobulin (Ig) secretion, and secretion of antibody to SRBC were determined. Lymphocytes cultured allogeneically at 5 X 10(6) cells per well in 24-well plates with SRBC and PCA (PWM, 1/10,000 dilution or LPS, 5 micrograms/ml) fused with the greatest efficiency. PWM-stimulated cultures fused on day 6 of in vitro stimulation yielded the greatest number of wells containing Ig-secreting hybridomas, whereas LPS-stimulated cultures produced more wells with hybridomas secreting Ig when fused on day 7 of stimulation. In vitro stimulation conducted in tissue culture flasks produced fewer wells with hybrid cell growth and Ig secretion. A fusion ratio of 1:1 (lymphocyte:WI-L2729HF2) yielded the most hybrid formation. Ig levels in wells with hybrid cell growth varied greatly, from less than 10 ng/ml up to 30 micrograms/ml. Ig of IgG, IgA, and IgM isotypes were produced. The addition of lymphocyte conditioned medium to in vitro immunization cultures does not improve the yield of Ig-secreting hybridomas. Hybrid cells secreting antibody to SRBC were cloned and analyzed for HLA specificities and chromosome numbers. Additionally hybridoma cells were formed after in vitro stimulation of peripheral blood lymphocytes (PBL) of normal individuals by fusion of the stimulated lymphocytes with WI-L2-729HF2. Hybridoma formation with PBL was less successful, but hybridoma cells secreting antibody to SRBC after in vitro stimulation were isolated.
Production of interleukin 2, interleukin 3, and interferon by mouse T lymphocyte clones of Lyt-2+ and -2- phenotype.
Authors: Guerne P A , Piguet P F , Vassalli P
Published: December 2022

T lymphocyte clones were derived by stimulation of positively selected Lyt-2+ and Lyt-2- lymphocytes with Con A in an Interleukin 2 (IL 2)-enriched medium under conditions of limiting dilution. Forty clones were expanded and tested, after activation by Con A, for the production of IL 2, IL 3, and interferon (IFN). Thirteen Lyt-2- clones were all co-producers of IL 2 and IL 3, and 10/13 were also producers of IFN. Twenty-seven Lyt-2+ clones were much more heterogeneous, 13 being IL 2 and IL 3 nonproducers, whereas 14 produced variable and poorly correlated amounts of IL 2 and IL 3. Three Lyt-2+ clones were observed to produce IL 2 or IL 3 alone. The majority of the Lyt-2- (10/13) and of the Lyt 2+ (21/27) clones were also producers of IFN. Exogenous IL 2 added during the activation of the Lyt-2+ by Con A did not enhance IL 3 production, whereas it did enhance IFN production by some but not all Lyt-2+ clones. Thus, among the T lymphocytes of the Lyt-2+ and -2- phenotypes there are cells capable of releasing IL 2, IL 3, and IFN. This supports the concept that these phenotypes are associated with antigen recognition rather than with cell functions, but it is apparent that the functional capacities of individual T cells, if accurately represented by their clonal progeny, are far from uniform.
Effect of fibronectin fragments on macrophage phagocytosis of gelatinized particles.
Authors: Rourke F J , Blumenstock F A , Kaplan J E
Published: December 2022

Rat plasma fibronectin enhances the binding and ingestion of gelatin-coated, formalin-fixed, or tanned sheep erythrocytes by elicited rat peritoneal macrophages. Fibronectin binding to the gelatinized erythrocytes is required for this enhancement, because macrophages preferentially recognize the surface bound molecule. This enhancement of particle uptake by fibronectin required the presence of a renewable, trypsin-sensitive component(s) on the macrophage surface (fibronectin receptor). When subjected to plasminolysis for 3 hr, fibronectin was degraded into gelatin-binding fragments (170 to 210 kd) and a 25-kd nongelatin binding fragment. The 170 to 210 kd gelatin binding fragments retained uptake-enhancing activity but were less active on a weight and molar basis than intact, dimeric fibronectin. The nongelatin binding 25 kd fragment alone did not enhance uptake. These results indicate that the sites for interaction with both the gelatinized erythrocyte surface and macrophages are retained on 170 to 210 kd fragments. However, the fibronectin dimeric structure is required for maximal expression of opsonic activity.
Separation of the immune response genes for LDH-B and MOPC-173. II. Description of an immune response defect in B10.ASR7.
Authors: Gutmann D H , Niederhuber J E
Published: December 2022

By using the intra-I region recombinant mouse strain B10.ASR7 (H-2as3), the immune response (Ir) genes for LDH-B and MOPC-173 were genetically and serologically separated, as assayed by T cell proliferation. Previous work demonstrated that H-2s and H-2b strains respond to LDH-B and MOPC-173 whereas H-2a and H-2k strains failed to respond due to haplotype-specific suppression of I-Ak molecule-activated T helper cells by I-Ek molecule-activated T suppressor cells. In the experiments reported here, B10.ASR7 mice, which lack I-Ek expression, mounted a significant T cell proliferative response to LDH-B but not to MOPC-173. Separation of the Ia determinants used in restricting these two antigen responses was further confirmed when pretreatment of B10.S(9R) (A beta sA alpha sE beta sE alpha k) macrophages with A.TL anti-B10.HTT serum (anti-As beta Es beta Js) adsorbed with B10.ASR7 spleen cells blocked the MOPC-173 response but not the LDH-B response. Unadsorbed serum blocked both antigen responses. The B10.ASR7 E beta allele was determined to be s due to the ability of (A.TL X B10.ASR7)F1 hybrids to mount a T cell proliferative response to the terpolymer GLPhe. Monoclonal antibody blocking of the B10.ASR7 T cell proliferative response to LDH-B demonstrated that the Ia.2 and Ia.17, and not the Ia.15 epitopes are spatially related to the Ia epitopes involved in the restriction of the B10.ASR7 LDH-B T cell proliferative response. In addition, B10.ASR7 helper T cells generated in response to LDH-B were suppressed in a haplotype-specific manner by I-Ek molecule-restricted suppressor T cells in that (A.TL X B10.ASR7)F1 hybrids failed to respond to LDH-B. This nonresponsiveness was eliminated by treatment with monoclonal antibodies directed against the I-Ek molecule. These results suggest the possibility that the immune response defect in B10.ASR7 could be related to the site of recombination.
Physiologic inactivation of fluid phase C3b: isolation and structural analysis of C3c, C3d,g (alpha 2D), and C3g.
Authors: Davis A E , Harrison R A , Lachmann P J
Published: December 2022

The fragments that result from the inactivation of C3b have not been completely characterized. Initial inactivation is catalyzed by the protease factor I, which, in the presence of its cofactor (factor H), cleaves two peptide bonds in the alpha'-chain of C3b. This results in the release of a small peptide (C3f, Mr 3000) from iC3b, which consists of the C3 beta chain covalently bonded to two alpha'-chain-derived peptides (Mr 68,000 and Mr 43,000). Surface-bound iC3b is cleaved at a third site by factor I to produce C3c and C3d,g (or alpha 2D). The factor I cofactor for this cleavage is the C3b receptor that is present on erythrocyte and leukocyte membranes. This report describes the isolation and initial structural characterization of C3c and C3d,g generated in whole blood after complement activation with cobra venom factor. These fragments were compared with the C3 fragments isolated from the serum and plasma of a patient with complement activation in vivo. The fragments were isolated with two solid phase monoclonal antibodies, one of which recognizes a determinant on C3g (clone 9) and one of which recognizes a determinant on C3c (clone 4). C3c isolated from normal blood showed three polypeptides that had apparent m.w. of 75,000, 43,000, and 27,000. The C3d,g consisted of a single polypeptide chain with a m.w. of 40,000. Amino terminal sequence analysis showed that the Mr 27,000 peptide from C3c is derived from the amino terminal portion of the alpha'-chain of C3b, whereas the Mr 43,000 peptide is derived from the carboxy terminus of the same chain. Amino terminal sequence analysis showed also that C3g is derived from the amino terminus of C3d,g. The C3 fragments isolated from a patient with partial lipodystrophy, nephritic factor activity, low serum C3 levels, and circulating C3 cleavage products showed a more complicated pattern on SDS-PAGE. The fragment isolated with clone 9 had an apparent m.w. of 40,000, identical to C3d,g generated in vitro, and it had the same amino terminal sequence as C3d,g generated in vitro. The eluate from insolubilized clone 4, however, showed prominent bands with Mr of 75,000, 56,000, 43,000, and 27,000, together with a triple-banded pattern at 68,000 and a minor band at 80,000. This eluate thus appears to contain C3c, and iC3b or an iC3b-like product. The origin of the Mr 56,000 and Mr 80,000 peptides have not yet been determined. These studies, with previous data, definitively order the C3c and C3d,g peptides in the alpha-chain of C3.(ABSTRACT TRUNCATED AT 400 WORDS)
Cellular origins and target cells of immunoregulatory factors in mice with disseminated histoplasmosis.
Authors: Deepe G S , Watson S R , Bullock W E
Published: December 2022

Disseminated infection with Histoplasma capsulatum stimulates the production of a suppressor factor (SF-H) by spleen cells from C3H/HeJ mice and a helper factor (HF-H) by spleen cells from C57BL/6 mice. In the present study these disparate immunoregulatory factors were analyzed in detail with regard to: a) the surface phenotype of the cells that produce SF-H and HF-H; b) the role of accessory cells in the production of these factors; and c) the surface phenotype of the target cells activated by SF-H and HF-H. Treatment of spleen cells from Histoplasma-infected C3H/HeJ mice with anti-Thy-1.2 plus complement (C) or with anti-Ly-2 plus C or with anti-I-Jk antiserum plus C abolished production of SF-H. Conversely, generation of HF-H by spleen cells from C57BL/6 mice was abrogated by treatment with either anti-Thy-1.2 plus C or with anti-Ly-1 plus C. Thus, a Thy-1.2+, Ly-2+, I-J+ T cell releases SF-H, and a Thy-1.2+, Ly-1+ T cell secretes HF-H. Production of SF-H and HF-H by splenic T cells was reduced markedly by depletion of macrophages (M phi); readdition of 1% syngeneic, plastic-adherent splenocytes from normal or infected mice to M phi-depleted, splenic T cell cultures of either strain restored the capacity to generate immunoregulatory factors. Furthermore, adherent splenocytes from normal or infected mice liberated a factor or factors that enhanced production of both SF-H and HF-H. Kinetic studies demonstrated that activation of normal spleen cells required at least 48 hr of exposure to SF-H or HF-H. Both factors failed to activate splenocytes pretreated with anti-Thy-1.2 plus C. Spleen cells from C3H/HeJ mice depleted of Ly-1+, Ly-2+, or I-J+ cells and exposed to SF-H did not demonstrate suppressor activity, whereas Ly-1-depleted splenocytes from C57BL/6 mice exposed to HF-H failed to exert helper activity. Therefore, the target of SF-H is a Thy-1.2+, Ly-1+2+, I-J+ T cell, and the target of HF-H is a Thy-1.2+, Ly-1+ T cell.
Modulation of Schistosoma mansoni egg-induced granuloma formation. III. Evidence for an anti-idiotypic, I-J-positive, I-J-restricted, soluble T suppressor factor.
Authors: Abe T , Colley D G
Published: December 2022

Modulation of pathogenic egg-induced hepatic granuloma formation in chronically Schistosoma mansoni-infected mice is an immunoregulatory process. Adoptive transfer and in vitro studies have demonstrated that this suppression involves various T lymphocyte circuitries, and the participation of soluble suppressor factors has recently been noted in these systems. The present study has partially characterized a soluble suppressive activity extracted from the thymus glands of chronically infected mice (SmTsF) that modulates granuloma formation in acutely infected mice. The suppressive effect of SmTsF could be administered by multiple i.v. injections or by slow release from osmotic minipumps implanted i.p. Homologous and reciprocal transfers of SmTsF prepared from B10.A(3R) and B10.A(5R) donors indicated that SmTsF-induced suppression required homology between the donor and recipient at the I-J subregion of the major histocompatibility complex. Furthermore, the use of immunoabsorbents prepared with anti-I-Jk and anti-I-Jb sera demonstrated that CBA/J (H-2k) SmTsF was retained by, and could be recovered from, anti-I-Jk insoluble columns, but was unaffected by parallel treatment with anti-I-Jb sera. Subsequent immunoabsorbent studies showed that SmTsF did not bind to soluble egg antigenic (SEA) columns, and thus demonstrated a lack of idiotype, anti-antigen activity. However, columns prepared by using anti-SEA IgG from chronically infected syngeneic mice retained SmTsF suppressive activity, and it could be recovered by alkaline elution. These data are compatible with an interpretation that the suppressive activity expressed anti-idiotypic reactivity. Thus a thymus extract obtained from chronic, modulated, S. mansoni-infected mice can induce granuloma suppression in acutely infected mice. This activity is associated with an I-J determinant-bearing, possibly anti-idiotypic moiety or moieties. These observations further implicate some of the Ts cascades reported in other systems in the regulation of cell-mediated pathogenesis in chronic experimental schistosomiasis.
Monoclonal antibody SN2 defining a human T cell leukemia-associated cell surface glycoprotein.
Authors: Seon B K , Negoro S , Barcos M P , Tebbi C K , Chervinsky D , Fukukawa T
Published: December 2022

We generated a monoclonal antibody, termed SN2, which defines a human T cell leukemia-associated cell surface glycoprotein, GP37, with an approximate m.w. of 37,000. This antibody was generated by using a human leukemia antigen preparation. The reactivity and specificity of SN2 were characterized by a sensitive radioimmunoassay against a variety of cultured and uncultured human cells. In selected cases, the cell specimens were tested further by indirect immunofluorescence staining. Among the various cultured malignant and nonmalignant human cell lines tested, SN2 reacted only with leukemic T cell lines, with one exception. It reacted with 10 of 11 leukemic T cell lines tested; the 10 reactive cell lines are PEER, JM, MOLT-4, CCRF-CEM, CCRF-H-SB2, RPMI 8402, DND-41, HPB-ALL, SKW-3, and HPB-MLT; the unreactive line was HUT 78. The reactive cell lines were derived from patients either with T cell-type acute lymphoblastic leukemia (the first eight cell lines), with T cell chronic lymphocytic leukemia (SKW-3), or with Japanese adult T cell leukemia-lymphoma (HPB-MLT). The unreactive cell line, HUT 78, was from a patient with Sezary syndrome. Results consistent with the above were obtained from studies in which uncultured malignant cell specimens from different cancer patients were tested against SN2; SN2 reacted only with T leukemia cells. Among various uncultured normal cell specimens tested, SN2 did not react with thymocytes, bone marrow cells, peripheral blood lymphocytes containing B and T cells, purified T cells, monocytes, granulocytes, or erythrocytes. It did, however, react with platelets.
Molecular signals in antigen presentation. II. Activation of cytolytic cells in vitro after ultraviolet radiation or combined gamma and ultraviolet radiation treatment of antigen-presenting cells.
Authors: Granstein R D , Tominaga A , Mizel S B , Parrish J A , Greene M I
Published: December 2022

Murine low-density spleen cells have potent antigen-presenting ability in a hapten-specific cytolytic T lymphocyte (CTL) system using the hapten azobenzenearsonate (ABA). Exposure of these cells to 0.33 KJ/m2 of ultraviolet radiation (UVR) after coupling to hapten results in markedly inhibited antigen-presenting function that can be substantially corrected or bypassed by interleukin 1 (IL 1). These results have been interpreted to reflect an inhibition of Lyt-1+ T cell activation by UVR-treated APC. Treatment of these cells sequentially with 1500 rad of gamma-radiation (GR) prior to hapten coupling, followed by 0.33 KJ/m2 of UVR radiation after coupling, results in an antigen-presenting defect only minimally improved by IL 1. However, partially purified interleukin 2 (IL 2) can completely bypass or correct this defect. Thus, combined GR and UVR induces a different or more profound defect in APC function when compared to UVR alone. However, these cells do provide a signal(s) other than hapten necessary for CTL activation because ABA-coupled high density spleen cells do not activate CTL cells, even with the addition of IL 2. Fluorescence-activated cell sorter analysis demonstrates that exposure of these low density spleen cells to GR or UVR results in decreased I-A antigen expression at 24 hr than either alone. The addition of nonhapten-coupled low-density APC partially reconstitutes the ability of combined GR/UVR-treated LD-APC to present antigen, and this effect is enhanced by the administration of exogenous IL 1. This occurs despite a lack of significant accessory cell activity by the LD-APC for the ABA hapten, and indicates that combined GR/UVR-treatment of APC is not functionally equivalent to completely removing them.