bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for covert cellular interaction sites expressed by activated T lymphocytes.
Authors: Thomas D W , Hoffman M D
Published: December 2022

In this report we examined the effects of trypsin treatment of immune guinea pig T lymphocytes on their binding to syngeneic macrophages. Immune T cells positively selected by culture for 1 wk with antigen and treated with trypsin showed dramatic binding to macrophages in the absence of additional antigen. This binding was rapid and greatest at 1 hr after the addition of trypsin-treated T cells to macrophages. Thereafter, the bound T cells dissociated from the macrophages and did not rebind. Dissociation of macrophage-bound T cells appeared to be an active process that could be inhibited by blocking T cell protein synthesis. Macrophage binding by trypsin-treated T cells was dependent on prior in vitro stimulation; similar treatment of immune T cells taken directly from the animal did not augment binding, and treatment of T cells cultured without antigen resulted in only modest binding. In addition, the trypsin-induced binding phenomenon showed elements of genetic restrictions. Trypsin treatment of immune T cells selected by culture for 1 wk with antigen resulted in preferential binding to syngeneic macrophages. In addition, immune F1 T cells selected by culture with antigen-pulsed parental macrophages preferentially bound to macrophages of the haplotype used for selection. Evidence is presented suggesting that the trypsin-induced binding was not simply due to antigen carry-over from the trypsinized lymphocytes and favoring the possibility that this binding may be antigen-independent. These results suggest that antigen-activated T cells express covert interaction sites for macrophages and that these sites are actively covered-up by other membrane components.
The antigens of pigeon breeder's disease. VII. Isoelectric focusing studies on unfractionated pigeon dropping extract.
Authors: McCormick D J , Fredricks W W , Tebo T H , Calvanico N J
Published: December 2022

Isoelectric focusing (IEF) studies on pigeon dropping extracts (PDE) revealed that it is a complex mixture of components that are acidic in nature. Chromatographically purified antigens PDEB, PDE1, and PDE3 showed multiple bands in IEF, indicating a microheterogeneity of these components, with peak concentrations focusing at pH 6.1, 4.6, and 3.8, respectively. The isoelectric points are compatible with the chromatographic behavior of these antigens on DEAE-cellulose. Crossed immunoelectrofocusing (CRIF) resolved PDE into seven precipitin lines with rabbit antiserum in a pH gradient from 3.5 to 9.5, and into nine precipitable components in a pH gradient from 2.5 to 7.0. The complexity of this antigen source appears to reside in the heterogeneity of immunologically related antigens. A comparison of pigeon breeder's sera by CRIF of PDE revealed a qualitative difference in precipitation patterns obtained with symptomatic and asymptomatic individuals. Sera from all (eight out of eight) of the symptomatic breeders tested precipitated an unidentified component of PDE, whereas none (zero out of four) of the sera from asymptomatic breeders detected this antigen. These results suggest that CRIF of PDE is useful as a diagnostic tool and that some specific component of PDE may be involved in the pathogenesis of the disease.
Regulation of T15 idiotype dominance. I. Mice expressing the xid immune defect provide normal help to T15+ B cell precursors.
Authors: Kenny J J , Wicker L S , Guelde G , Scher I
Published: December 2022

The immune response to phosphocholine (PC) in many strains of mice is dominated by the T15 idiotype family of anti-PC antibodies. By introducing the CBA/N X-linked immune defect (xid gene) into these mice, one profoundly alters their ability to make a T15-predominant, IgM anti-PC response. This loss of T15 dominance in mice expressing the xid gene is not due to the presence of suppressor T cells or the lack of T15 idiotype-specific helper cells in these mice. Thus, one can reconstitute a T15 idiotype-dominant response in immune defective mice with B cells from normal mice, and in adoptive transfer assays the primed T helper cells from immune-defective mice provide qualitatively the same help to normal B cells as the T helper cells from normal mice. T15 idiotype dominance appears to be controlled by the expression and activation of Lyb-5+ PC-specific B cells. Thus, the majority of T15+ B cell precursors are restricted to this B cell subset, whereas the Lyb-5- B cell subset contains predominantly T15-, anti-PC B cell precursors, which produce mainly IgG antibodies after activation by PC-containing antigens.
Immunologic memory to phosphorylcholine III. IgM includes a fine specificity population distinct from TEPC 15.
Authors: Chang S P , Brown M , Rittenberg M B
Published: December 2022

Group I and group II variable regions expressed in the PC-KLH memory response may originate from distinct germ-line genes or group II may arise by somatic mutation of group I (TEPC 15) germ-line genes. A current version of the somatic mutation hypothesis proposes that somatic mutation is activated by the class switch from IgM to IgG or IgA. If group II results from somatic mutation during class switching, group II IgM antibodies would not exist. This prediction was tested in the present experiments. Group I and group II antibodies were separated from whole serum by affinity chromatography on PC-Sepharose. An ELISA was used to characterize the fine specificity and idiotype of the isolated antibody populations. Group I antibodies were inhibited by both PC and NPPC haptens and were T15 idiotype-positive. Group II antibodies were inhibited appreciably only by NPPC and were negative for the T15 idiotype. The purified group II antibodies contained a significant IgM component, and high levels of group II IgM were detected during the early secondary response to PC-KLH. These observations are inconsistent with the hypothesis that group II originates by somatic mutation activated by the class switch. These results strongly suggest that one or more of the germ-line genes (V, D, or J) of groups I and II are different. Alternatively, if group II antibodies arise from group I by somatic mutation, this mutation must occur before class switching.
Amphotericin B alters the affinity and functional activity of the oligopeptide chemotactic factor receptor on human polymorphonuclear leukocytes.
Authors: Lohr K M , Snyderman R
Published: December 2022

Leukocyte chemotaxis is initiated by the binding of chemotactic factors to specific, high-affinity receptors. Amphotericin B, a polyene antibiotic that binds to membrane cholesterol, inhibits human neutrophil (PMN) chemotaxis. We examined the effects of this drug on PMN functions mediated by the oligopeptide chemotactic factor receptor. The antibiotic irreversibly inhibited chemotaxis and depressed the binding of the radiolabeled chemoattractant, fMet-Leu-[3H]Phe, to its receptor without affecting the receptor's specificity. The drug lowered the binding affinity of the receptor by up to fivefold and slightly increased its number. Doses of amphotericin B that depressed receptor affinity and inhibited chemotaxis did not diminish lysosomal enzyme secretion or superoxide anion production. Nystatin, a less potent polyene antibiotic, also diminished chemotactic factor binding, but to a lesser degree than amphotericin B did. A chemically unrelated antifungal agent had no effect on either binding or chemotaxis. Thus, pharmacologic manipulation can alter the affinity of the chemotactic factor receptor on human PMN; this alteration is associated with a change in receptor function. The data suggest that receptor affinity regulates or at least reflects its functional state, and that the transduction mechanisms for various biologic responses mediated by the chemoattractant receptor are heterogeneous. By pharmacologic alterations of receptor affinity, one may be able to modulate specific biologic responses elicited by chemoattractant receptor-ligand interactions.
The formylpeptide chemotactic receptor on rabbit peritoneal neutrophils. I. Evidence for two binding sites with different affinities.
Authors: Mackin W M , Huang C K , Becker E L
Published: December 2022

F Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe binding to rabbit peritoneal neutrophils and purified membranes were measured at 4 degrees C silicone oil centrifugation assays, and the results were analyzed by the LIGAND computer program, which permits analysis of ligand binding to multiple classes of binding sites. LIGAND analysis of peptide binding to intact neutrophil indicated that both f Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe detected two population of binding sites. The apparent Kd values for f Met-Leu-[3H]Phe binding were 1.6 +/- 1.0 X 10(-9) M and 2.2 +/- 0.9 X 10(-8) M, respectively, and 3.1 +/- 0.2 X 10(-9) M and 1.2 +/- 0.6 X 10(-7) M for f Nle-Leu-[3H]Phe. Furthermore, the higher affinity sites detected on whole cells comprised approximately 15 to 30% of the total sites. Two populations of binding sites were also detected on purified neutrophil plasma membranes by both radiolabeled chemotactic peptides. LIGAND analysis of peptide binding to purified membranes yielded apparent Kd values of 5.0 +/- 2.5 X 10(-10) M and 4.8 +/- 0.6 X 10(-8) M for f Met-Leu-[3H]Phe binding, and 4.7 +/- 4.2 X 10(-10) M and 3.0 +/- 1.3 X 10(-8) M for f Nle-Leu-[3H]Phe. The percentage of higher affinity sites detected by f Met-Leu-[3H]Phe and f Nle-Leu-[3H]Phe on purified membranes were 1 to 5% of the total sites detected. These data are consistent either with the existence of two independent binding sites for formylpeptides on rabbit neutrophils or receptor negative cooperativity.
Specific binding of phospholipid platelet-activating factor by human platelets.
Authors: Valone F H , Coles E , Reinhold V R , Goetzl E J
Published: December 2022

The binding of the phospholipid platelet-activating factor 1-O-hexadecyl-2-acetyl-sn-glycero-3-phosphorylcholine (AGEPC) to washed human platelets was more than 80% complete within 2 min, which coincided with the time of initiation of platelet aggregation by AGEPC. Scatchard plot analysis of the binding of [3H]AGEPC to platelets without and with an excess of unlabeled AGEPC revealed two distinct types of binding sites. One platelet site for AGEPC exhibited a high affinity (KD = 37 +/- 13 nM, mean +/- SD), was saturable, and had a low maximal capacity of 1399 +/- 498 (mean +/- SD) molecules of AGEPC/platelet. The other platelet site demonstrated a nearly infinite binding capacity, consistent with nonreceptor uptake of AGEPC into cellular structures. The specificity of the high-affinity binding site for AGEPC was assessed by comparing the capacity of several analogues of AGEPC to inhibit the binding of [3H]AGEPC to platelets and to induce platelet aggregation. An ether linkage in position 1, a short-chain fatty acid in position 2, and a choline moiety in the polar head group proved to be critical both for the binding of [3H]AGEPC to platelets and for the initiation of platelet aggregation. Exposure of platelets to AGEPC for 5 min at 37 degrees C functionally deactivated the exposed platelets to subsequent stimulation by AGEPC, as assessed by diminished aggregation, and concomitantly reduced the specific binding of [3H]AGEPC. Evaluation of the time course of the events of deactivation revealed the loss of an aggregation response to AGEPC after 90 sec at 37 degrees C, despite the retention of up to 50% of the specific binding sites for AGEPC.
Macrophages in resistance to rickettsial infections: genetic analysis of susceptibility to lethal effects of Rickettsia akari infection and development of activated, cytotoxic macrophages in A and B10.a mice.
Authors: Meltzer M S , Nacy C A , Stevenson M M , Skamene E
Published: December 2022

Susceptibility to lethal effects of Rickettsia akari varies among mouse strains. Although most strains are resistant, A/J mice are extremely sensitive (10,000-fold difference in LD50 between resistant and sensitive strains). In contrast to most strains of mice, A/J mice also fail to develop activated, tumoricidal macrophages after any of several in vivo or in vitro treatments. Are susceptibility to R. akari and inability to develop activated, cytotoxic macrophages in the A/J strain causally related traits? Genetic analysis of macrophage tumoricidal activity in responsive B10.A and nonresponsive A mice suggested this trait was controlled by a single, autosomal, dominant gene. Among (F1 X A) backcross mice, the traits for defective macrophage cytotoxicity and abnormal accumulation of macrophages during inflammation (a response controlled by a single, autosomal, dominant gene) segregated independently. Genetic analysis of resistance to R. akari in sensitive A and resistant B10.A mice suggested this trait was also controlled by a single, autosomal, dominant gene. Among (F1 X A) backcross mice, however, traits for defective macrophage cytotoxicity and R. akari resistance segregated independently. Thus, development of activated tumoricidal macrophages, accumulation of macrophages at sites of inflammation, and resistance to the lethal effects of R. akari in A and B10.A mice were each controlled by single, autosomal, dominant, non-H-2-linked genes. However, genes for control of macrophage activation for tumoricidal activity and for resistance to lethal effects of R. akari infection are distinct.
B cell expression of endogenous retroviral envelope antigen: biochemical and immunofluorescence characterization.
Authors: Ewert D L , Halpern M S
Published: December 2022

Plasma cells of line 15I5 chickens have been shown to express antigen that is cross-reactive with endogenous retroviral envelope glycoprotein. In the present study, we isolated this antigen and showed that it is structurally homologous to envelope glycoprotein. As assayed by immunofluorescence, most, if not all, of the plasma cell-associated envelope glycoprotein is present on the cell surface although it is not bound to surface Ig or Ia antigen. Although envelope antigen was not detectable by immunofluorescence on the surface of 15I5 bursal cells or peripheral B lymphocytes, immunochemical assays established that retroviral envelope glycoprotein is present on the surface of the bursal cells but at much lower levels than on the surface of the plasma cells. Both bursal and plasma cells synthesize envelope glycoprotein. These findings indicate that an increased level of expression of endogenous retroviral envelope glycoprotein is a concomitant of the differentiation of B lymphocytes to plasma cells in 15I5 chickens.
MLR-specific suppressor T lymphocytes in man. II. Functional and membrane characteristics of a specific MLR suppressor subpopulation.
Authors: Fabio G , Scorza-Smeraldi R , Bonara P , Sabbadini Villa M G , Vanoli M , Calabi F , Zanussi C
Published: December 2022

Human MLR-specific suppressor T lymphocytes were induced by in vitro cultivation of human peripheral T lymphocytes in the presence of soluble HLA-DR alloantigens isolated from normal serum. The suppression is specific in that responder cells autologous to the suppressor cells respond to allogeneic stimulating cells that express the same HLA-DR specificity as that recovered from serum and used to induce the suppressor cells. This antigen-specific suppressor T cell population could be divided into suppressor and non-suppressor subpopulations as a function of adherence to a 6MB Sepharose immunoadsorbent coated with the inducing HLA-DR soluble antigen. As a consequence of activation by soluble DR antigen, the suppressor T lymphocyte population as well as the column-enriched suppressor subpopulation express new membrane specificities that can be recognized by antisera from pluriparous women. The specificities that are recognized are not found on autologous, unstimulated B and T cells, nor do they appear to recognize conventional HLA-A,B,C or DR determinants.
Evidence for an IgD homologue on chicken lymphocytes.
Authors: Chen C L , Lehmeyer J E , Cooper M D
Published: December 2022

Chicken lymphocyte membrane immunoglobulins (Ig), were precipitated with mouse monoclonal antibodies specific for heavy and light chain isotypes and analyzed by polyacrylamide gel electrophoresis. Very little or no membrane-bound IgG and IgA was detected. After sequential precipitation and removal of IgM reactive with any of three monoclonal anti-mu antibodies, anti-light chain antibody precipitated residual Ig with a relative electrophoretic mobility similar to that of IgM. Under reducing conditions, these surface Ig molecules had a heavy chain that appeared slightly larger (approximately 81,000 daltons) than mu-chain (approximately 79,000 daltons), and light chains of approximately 25,000 daltons. Complete clearance of membrane-bound IgM reactive with an anti-mu allotype antiserum left similar molecules precipitate by monoclonal anti-light chain antibody. These non-IgM molecules could be detected on the surface of lymphocytes from blood, spleen, bursa and the B cell line RAV-1, but not from thymus or blood from an agammaglobulinemic chicken. After capping of B cell surface IgM with anti-mu, immunofluorescent staining with anti-light chain antibody revealed residual Ig molecules disturbed across the surface of more than 90% of the IgM-bearing cells. The data suggest the existence of an avian homologue of mammalian IgD. Affinity-purified goat anti-mu antibodies and a fourth monoclonal anti-mu antibody reacted with both IgM and the putative IgD molecules, which suggests that the IgD homologue shares at least one common determinant with chicken IgM.
Haplotype-specific suppression of T cell response to lactate dehydrogenase B in (responder x nonresponder)F1 mice.
Authors: Nagy Z A , Baxevanis C N , Klein J
Published: December 2022

Mouse strains that express the Ek (Ek beta E-1k alpha) molecule are nonresponders (NR) to the enzyme lactate dehydrogenase B (LDHB) in terms of T cell proliferation. Nonresponsiveness is caused by T suppressor (Ts) cells recognizing LDHB in the context of Ek molecules on the antigen-presenting cells. The data presented here demonstrate that the Ek-restricted Ts cells function in (R x NR)F1 mice in a remarkable haplotype-specific fashion: they selectively interfere with the Ak (ANR)-restricted response, and do not affect the response channeled through the A molecules of the responder parent. This haplotype-specificity of suppression provides an explanation of the dominance of responsiveness in (R x NR)F1 mice.
Evidence that retrovirus expression in mouse spleen cells results from B cell differentiation.
Authors: Alberto B P , Callahan L F , Pincus T
Published: December 2022

Induction of endogenous retroviral proteins from spleen cells exposed to lipopolysaccharide (LPS) was studied in individual cells in relation to polyclonal B cell differentiation. After 4 days of culture with LPS, 30 to 40% of cells in many mouse strains stained for cytoplasmic mu heavy chains. A smaller number of lymphoid cells stained for cytoplasmic viral proteins, with a characteristic proportion for each strain, including 21% of lymphoid cells of AKR mice, 125 of DBA/2 cells, 7% of C57BL/6 cells, and 2% of BALB/c cells. More than 75% of retrovirus-positive cells were also mu-chain-positive in all strains. Cells from young (3- to 8-week-old) and old (43- to 46-week-old) mice of several strains showed similar proportions of retrovirus-positive cells typical for each strain, without differences at different ages. The induction of retrovirus proteins, as well as mu-chains, by LPS was markedly inhibited by goat anti-mu-chain antibody added to the culture medium (10 to 100 micrograms/ml). These data suggest that retrovirus induction may result from B cell differentiation.
Bacteriophage Purified with Lipoids
Authors: Weiss Emil
Published: December 2022

Bacterial proteins in a phagic fluid can be precipitated by lipoids in a proper concentration. This purified bacteriophage produces only antilysins, is nontoxic, has no prophylactic value and is very useful for therapeutic purposes.
A monoclonal antibody from infected mice to a Schistosoma mansoni egg proteinase.
Authors: Dresden M H , Sung C K , Deelder A M
Published: December 2022

A number of monoclonal antibodies were obtained by fusion of SP2/0 myeloma cells and spleen lymphocytes from mice infected with Schistosoma mansoni. These antibodies were tested for their ability to inhibit acidic, thiol-dependent proteinases previously isolated from Schistosoma mansoni eggs and adult worms. One of the monoclonal antibodies isolated inhibits egg proteinase activity measured in vitro with the use of a low m.w. synthetic substrate. This antibody, which is an IgG1 isotype, does not appreciably inhibit an acidic, thiol-dependent proteinase obtained from the adult stage of Schistosoma mansoni. Immunocytochemical methods with the monoclonal antibody have been used to localize the egg proteinase within a set of "penetration" glands in the unhatched miracidium.
Cell surface phenotype of lymphoid cells from normal mice and mice treated with monoclonal anti-IgD from birth.
Authors: Skelly R R , Baine Y , Ahmed A , Xue B , Thorbecke G J
Published: December 2022

Mice treated from birth with mouse monoclonal anti-IgD antibodies develop low frequencies of B cells in the spleen, a small percentage of which express very low levels of sIgD on their cell surface and extremely low frequencies of B cells in their lymph nodes, lacking sIgD entirely. However, the splenic B cells are phenotypically mature in that a high percentage of these cells express Lyb-5, indicating that the expression of sIgD is not a prerequisite for the acquisition of a mature surface antigen repertoire of B cells. In contrast, a high density of sIgM on splenic B cells is expressed, which suggests a predominance of cells with the phenotype of immature B cells and/or activated B cells. Furthermore, the spleen cells from anti-IgD-treated mice lack cells that respond to in vitro stimulation by LPS with an increase in the density of their sIa.
Accumulation of guanine-cytosine-enriched low M.W. DNA fragments in lymphocytes of patients with systemic lupus erythematosus.
Authors: Sano H , Imokawa M , Steinberg A D , Morimoto C
Published: December 2022

The fate of the newly synthesized DNA in peripheral blood lymphocytes from patients with systemic lupus erythematosus was examined. After lymphocytes were stimulated with the mitogen phytohemagglutinin (PHA), DNA was pulse-labeled with radioactive thymidine and was analyzed by sucrose density gradient centrifugation. Two classes of DNA were identified; the main DNA fraction that migrated to the same position as that of control samples, and low m.w. DNA fragments. The low m.w. DNA fragments were further characterized: i) The amount of low m.w. fragments increased with increasing time of culture after PHA stimulation. ii) The guanine cytosine content was 47% of the total bases at 4 days after PHA stimulation. iii) The base composition was similar to that of DNA fragments isolated from DNA/anti-DNA antibody immune complexes. These results lead us to propose that guanine cytosine-enriched low m.w. DNA fragments accumulate in cells after nuclear DNA is degraded, and that these fragments may serve as a primary source of autoantigen for anti-DNA antibody production. In addition, they may interfere with normal cellular metabolism.
Cell surface antigens and other characteristics of T cells regulating the antibody response to type III pneumococcal polysaccharide.
Authors: Taylor C E , Amsbaugh D F , Stashak P W , Caldes G , Prescott B , Baker P J
Published: December 2022

The administration of a subimmunogenic dose of type III pneumococcal polysaccharide (SSS-III) produces an antigen-specific T cell-dependent phenomenon termed low-dose paralysis (immunologic unresponsiveness). This form of unresponsiveness can be transferred by spleen cells obtained 5 to 24 hr after priming, and the suppressive activity of the transferred cells is abolished by prior treatment with monoclonal anti-Lyt-2 and anti-I-J antibody in the presence of complement, indicating that suppression is mediated by a distinct subset of T cells (suppressor T cells). If primed spleen cells are transferred 24 to 72 hr after immunization with SSS-III, however, the resulting antibody response of immunized recipients is enhanced. Greater enhancement is noted when transferred cells, pretreated with monoclonal anti-Lyt-2 antibody plus complement to remove suppressor T cells, are used; such enhancement is attributed to amplifier T cells. These findings indicate suppressor T cells regulate the antibody response to SSS-III by influencing the expansion of SSS-III-specific clones of B cells as well as the expression of amplifier T cell activity; the latter causes B cells to proliferate further in response to SSS-III.
A human T cell-derived soluble factor able to suppress pokeweed mitogen-induced immunoglobulin production.
Authors: Warrington R J , Sauder P J , Olivier S L , Rutherford W J , Wilkins J A
Published: December 2022

Human peripheral blood T cells or Fc gamma receptor-positive T cells activated with the mitogen concanavalin A produce a soluble suppressor factor that inhibits polyclonal B cell immunoglobulin production. The factor suppresses pokeweed mitogen- (PWM) induced plaque-forming cells (PFC) by a noncytotoxic effect. It does not significantly affect lymphocyte proliferation, and exerts its effect independently of the presence of T cells. It is active in suppressing PFC when added from day 0 to day 3 of PWM cultures. Suppression is blocked by the addition of the monosaccharide L-rhamnose. The factor is stable at pH 2.0 and stable at 56 degrees C, but is inactivated by 2-mercaptoethanol. The approximate m.w. by gel filtration is 20,000 to 25,000. The above characteristics distinguish this suppressor factor from soluble immune suppressor supernatant, lymphotoxin, and gamma-interferon. They do not distinguish the suppressor factor from migration inhibition factor, which is also present in the supernatants.
Separation of three class II antigens from a homozygous human B cell line.
Authors: Shackelford D A , Lampson L A , Strominger J L
Published: December 2022

Three class II molecules were isolated from a homozygous DRw6 human B lymphoblastoid cell line using the monoclonal antibodies L243 (L203), L227, LKT 111, and Genox 3.53. Two of the antigens appeared to employ the same heavy chain but expressed different light chains. The two light chains were separated after denaturation using L227 and LKT 111. One or both of these two molecules carried the DRw6 and MT2 determinants. The third class II antigen expressed the DC1 determinant. It was composed of a heavy and light chain different from the DR-like antigen subunits. The antibodies L243, L227, and LKT 111 did not preclear the cell lysate of the DC1 antigen recognized by Genox 3.53. However, a xenoanti-DR serum immunoprecipitated both the DR-like and the DC1 antigens. Thus, in total, one cell line can express at least two class II heavy chains and three class II light chains. This observation was not unique to this cell line.
Organ-dependent differences in composition and function observed in hepatic and intestinal granulomas isolated from mice with Schistosomiasis mansoni.
Authors: Weinstock J V , Boros D L
Published: December 2022

In murine schistosomiasis mansoni, eggs deposited in the liver and intestines induce a cell-mediated granulomatous reaction. Previous studies have shown that maximal granuloma size differs in the liver and various segments of the gastrointestinal tract. The objective of this investigation was to isolate intestinal granulomas and to determine whether organ-dependent differences in cell composition and granuloma function exist. Intestinal granulomas representative of those in tissue were isolated by a combination of chemical and mechanical techniques. When dissociated by collagenase, these lesions yielded a viable heterogeneous population of inflammatory cells. Granulomas from the liver, colon, and ileum showed differences in cellular composition. Liver lesions contained the largest number of T and B lymphocytes, eosinophils, and mast cells whereas ileal granulomas comprised mostly macrophages. Immunofluorescence studies on frozen tissue sections revealed that T and B lymphocytes also displayed different patterns of distribution within granulomas from different tissues. In contrast to isolated cultured liver granulomas that produced MIF-like substances, isolated colonic and ileal granulomas had weak or no MIF activity. It thus appears that granuloma formation in various organs is influenced by local factors that could affect the ultimate resolution of the lesions.
In vivo development of cytolytic T lymphocytes (CTL) to hapten-altered self: MIs-disparate cells facilitate the response by neutralizing IL 2 inhibitor.
Authors: Gautam S C , Hilfiker M L , Battisto J R
Published: December 2022

Inability to develop CTL in vivo to hapten-altered self can be attributed in part to an inhibitor of interleukin 2 (IL 2) that is present in the serum of normal mice. We have shown earlier that hapten-specific CTL can be generated in C3H mice (H-2k, MIsc) provided CBA/J (H-2k MIsd) spleen cells are injected simultaneously with hapten-modified syngeneic spleen cells into the hind foot paws. In efforts to determine whether serum levels of the inhibitor of IL 2 are altered as a consequence of this successful immunization method, we have compared the activity of the inhibitor in serum at intervals after the injection of syngeneic spleen cells, CBA spleen cells, or TNP-C3H spleen cells alone or together with CBA spleen cells, by using a murine IL 2-dependent, cloned cytotoxic T cell line, CT-6. The results indicate that inhibitor was neutralized optimally 48 to 72 hr after injection of TNP-C3H spleen cells mixed with CBA/J spleen cells. The order in which neutralization occurred was as follows: TNP-C3H cells + CBA/J cells greater than CBA cells greater than TNP-C3H cells greater than normal C3H spleen cells. Furthermore, supernatants from cultures of C3H lymph node cells stimulated in vivo with CBA/J cells also contained IL 2 activity. Thus, injection of CBA/J cells with TNP-modified syngeneic spleen cells produces IL 2 in vivo in sufficient quantity to neutralize the activity of the inhibitor as well as to facilitate the maturation of pre-CTL into hapten-altered self-specific CTL.
Activation of lymphocytes by a thiol-derivatized nucleoside: characterization of cellular parameters and responsive subpopulations.
Authors: Goodman M G , Weigle W O
Published: December 2022

Lymphocyte activation, whether specific or nonspecific, is generally conceptualized as initiated by the binding of an activating ligand to a surface membrane receptor, followed by transduction of the signal across the cell membrane. In many situations several qualitatively distinct signals are required. We have recently described a new class of lymphocyte activator, the C8 bromine substituted guanine ribonucleosides, that traverse the cell membrane, bypassing classical triggering mechanism(s), apparently activating the lymphocyte at an intracellular site. However, the identity of the lymphocyte population(s) activated, as well as the nature of any cellular interactions involved in activation, has not been studied heretofore. The present experiments describe the cellular parameters of lymphocyte activation by a thiol substituted member of this class of activators, 8-mercaptoguanosine (8MGuo). Upon addition of this nucleoside derivative to cultures of murine spleen cells, a marked increase in [3H]TdR uptake and blast transformation ensues. Normal splenic B cells and spleen cells from congenitally athymic (nu/nu) mice are responsive to 8MGuo, whereas thymocytes and splenic T cells are not. Two subpopulations of B cells appear to be involved in the response to this nucleoside. The predominant one is a mature population that bears surface delta-chains, la antigens, C receptors, and (by indirect evidence) the Lyb3, 5, and 7 antigens. These cells also bear mu-chain and Fc receptors. In addition, a second, minor subpopulation of less mature cells that bear only mu-chain and Fc receptors also appears to be reactive to 8MGuo. The existence of this immature, reactive B cell subset was confirmed by observation of 8MGuo responsiveness in lymphocytes from 4-day-old mice whose cells do not yet exhibit these later-appearing markers. Accessory cells appear to play a minimal, if any, role in the 8MGuo response. These results establish two distinct B cell subpopulations as the major and minor cellular targets of C8-derivatized nucleosides, and suggest that the activation process results from a direct interaction between the nucleoside and target cell.
Idiotype shifts caused by neonatal tolerance to phosphorylcholine.
Authors: Quintáns J , Quan Z S
Published: December 2022

The injection of as little as 0.5 microgram phosphorylcholine-(PC) conjugated mouse immunoglobulin into BALB/c neonates within 48 hr of birth results in complete unresponsiveness to PC for 3 to 4 wk. Thereafter, anti-PC responses can be detected in tolerized animals, but these responses differ significantly from those of normal BALB/c mice. First, the magnitude of responsiveness does not approach normal levels even 9 mo after birth. Second, although the initial responses as tolerance is broken can be T15+, idiotypic dominance is not established; instead, a heterogeneous T15- population eventually emerges, which includes clones with higher and with lower avidity than T15. Unirradiated unresponsive mice will help transplanted normal B cells to produce T15+ responses to thymus-dependent PC antigens. The responses of animals recovered from tolerance are stable upon adoptive transfer. We have, moreover, found no evidence of either loss of idiotype-specific T cell help or generation of suppression. Therefore, neonatal exposure to PC tolerogen can effect profound, permanent changes in the antigen-specific B cell compartment independent of any influence on conventional T cell regulatory mechanisms.
Analysis of the B cell subpopulations influenced by allogeneic effect factor. I. MHC restricted enhancement of b cell responses to thymic-independent antigens, types 1 and 2, in normal and CBA/N mice.
Authors: Yaffe L J , Mond J J , Ahmed A , Scher I
Published: December 2022

The ability of allogeneic effect factor (AEF) to enhance the in vitro antibody responses of murine T cell-depleted B cell populations to thymic-dependent (TD) antigens also occurs with thymic-independent (TI) antigens. AEF significantly enhanced the in vitro antibody responses of normal T cell-depleted spleen cell populations to both 2,4,6-trinitophenylated Brucella abortus (TNP-BA), a TI type 1 antigen, and 2,4,6-trinitrophenylated aminoethylcarbamylmethyl-Ficoll (TNP-Ficoll), a TI type 2 antigen. The enhancing activity of AEF on TI responses was analyzed further in immune-defective CBA/N mice that possess only Lyb-5- B cells, and thus fail to respond to TNP-Ficoll, but do respond to TNP-BA and sheep red blood cells (SRBC). AEF was able to reconstitute the in vitro antibody responses of CBA/NT cell-depleted B cell populations to SRBC and to augment the T cell-depleted response to TNP-BA. These data suggest that AEF can enhance the activity of both Lyb-5+ as well as Lyb-5- B cells in their responses to both type 1 and type 2 TI antigens, and to the TD antigen SRBC. This enhancing effect of AEF on the TD and TI responses was MHC restricted for both Lyb-5+ as well as Lyb-5- spleen cells.
Functional differentiation in the genetic control of murine T lymphocyte responses to human fibrinopeptide B.
Authors: Peterson L B , Wilner G D , Thomas D W
Published: December 2022

The ability to generate proliferative and helper T lymphocyte responses in mice was compared by using the 14 amino acid peptide, human fibrinopeptide B (hFPB). Lymph node or peritoneal exudate T cells from mice immunized with hFPB were assessed for in vitro proliferation to soluble hFPB as determined by the uptake of 3H-thymidine. The T cell proliferative response to hFPB was found to be under MHC-linked Ir gene control; mice possessing the H-2a,k haplotypes were responders, whereas H-2b,d,q,s mice were nonresponders. The influence of non-H-2 genes on these responses was not investigated, so exclusive regulation by H-2 is provisional. The absence of a detectable lymph node and peritoneal exudate T cell proliferative response persisted in H-2b,d,q,s mice after immunization and boosting with several doses of hFPB. In addition, the capacity to produce a T cell proliferative response was inherited in an autosomal dominant manner and gene(s) controlling responsiveness to hFPB mapped to the I-A subregion of the H-2 complex. To measure peptide-specific helper T cell activity, an in vitro microculture assay in which hFPB-primed lymph node T cells and normal spleen B cells and macrophages were used was developed measuring anti-fluorescein isothiocyanate (FITC) IgM and IgG plaque-forming cell (PFC) responses after culture with FITC-conjugated peptide. Immunization of B10.BR, C57BL/10, B10.D2, and B6AF mice with hFPB primed for significant helper T cell activity as assessed by the ability to augment a primary in vitro IgM response to FITC. The normal B cell IgM responses were completely dependent on hFPB-primed T cells and required that hapten (FITC) and carrier (peptide) be linked. In addition, immunization with FITC-conjugated peptide elicited positive in vivo PFC responses to FITC in B10.BR and C57BL/10 mice, indicating similar genetic control of helper activity in both the intact animals and the in vitro microcultures. Thus, B10.BR mice show both T help and T proliferative responses to hFPB, whereas C57BL/10 mice show only T help and no T proliferative responses. In contrast to B10.BR mice, C3H and CBA mice immunized with hFPB were completely unresponsive when assayed for helper T cell activity in vitro despite their ability to generate positive lymph node T cell proliferative responses. These results indicate responsiveness to hFPB by T helper and proliferating cells is different and is under separate genetic control.
Genetic studies of autoimmunity in New Zealand mice. IV. Contribution of NZB and NZW genes to the spontaneous occurrence of retroviral gp70 immune complexes in (NZB X NZW)F1 hybrid and the correlation to renal disease.
Authors: Maruyama N , Furukawa F , Nakai Y , Sasaki Y , Ohta K , Ozaki S , Hirose S , Shirai T
Published: December 2022

We designed a polyethylene glycol precipitation method for detecting retroviral gp70-anti-gp70 immune complexes (gp70 IC) in the circulation of mice. To determine the genetic contribution of NZB and NZW strains to the spontaneous occurrence of circulating gp70 IC in NZB X NZW (B/W)F1 hybrids, we measured these IC in female NZB, NZW, (NZB X NZW)F1 (B/W F1), B/W F1 X NZW backcross, and B/W F1 X NZB backcross mice. The highest amounts of gp70 IC were found in B/W F1 and the lowest in NZW mice, and the order of the average serum levels was B/W F1, B/W F1 X NZB, B/W F1 X NZW, NZB, and NZW. Genetic analyses suggested that two major dominant genes, one from NZB and the other from NZW, are involved in the formation of these gp70 IC. A single dominant gene of NZB strain determines the formation of these IC, and the magnitude is to a great degree modified (intensified) by the NZW gene. In addition, statistical analyses of data obtained in the studies of B/W F1 X NZW backcrosses suggested the presence of one additional dominant NZB gene that also regulates the magnitude of gp70 IC formation in concert with the other NZB gene. Since all these New Zealand mice share a high serum level of free gp70, the formation of gp70 IC represents the gene action on the production of anti-gp70 antibodies. Linkage studies showed that both of the major NZB and NZW genes are loosely linked to the H-2 complex but not to either the Mup-1 or coat color gene loci examined. The incidence of proteinuria correlated well with the serum level of gp70 IC in both B/W F1 X NZW and B/W F1 X NZB backcross mice.
Activation of antibody-dependent cell-mediated cytotoxicity of human neutrophils and eosinophils by separate colony-stimulating factors.
Authors: Vadas M A , Nicola N A , Metcalf D
Published: December 2022

Semi-purified human colony-stimulating factors (CSF) powerfully enhanced the antibody-dependent cell-mediated cytotoxicity (ADCC) by metrizamide gradient-purified human neutrophils and eosinophils. The stimulation was observed on three different tumor targets, was rapid (less than 1 hr) in onset, and CSF-stimulated cells needed direct contact with targets for killing. A subspecies of human CSF, CSF-alpha, with eosinophil and granulocyte-macrophage (GM) colony-stimulating activity enhanced both eosinophil and neutrophil killing. In contrast, another subspecies of human CSF, CSF-beta, having only GM colony-stimulating activity, only enhanced neutrophil-mediated ADCC. These results support the notion that human CSF have two sites of action: i) the progenitor cell, where they stimulate a relatively slow process of differentiation, and ii) the mature cell, where they have a rapid action of increasing functional capacity. Furthermore, it seems the pattern of CSF receptors on progenitor cells is maintained throughout the lineage of such cells and serves to regulate the function of mature cells.
The extracellular region of light chains from human and murine MHC class II antigens consists of two domains.
Authors: Kaufman J F , Strominger J L
Published: December 2022

The experiments in this report, together with previous studies, demonstrate that the light chains of DR antigens are composed of four domains: two large extracellular domains (each with a disulfide loop), a short hydrophobic membrane-binding region, and a short hydrophilic carboxy-terminal domain. The amino-terminal extracellular domain is glycosylated and polymorphic and has no discernible sequence homology to immunoglobulin, whereas the relatively conserved carboxy-terminal extracellular domain bears striking homology to immunoglobulin. One chymotryptic and two tryptic cleavage sites lie in the second extracellular domain of the light chain of all native DR antigens. These three sites are found in loops at the same end of an immunoglobulin-like domain. The light chain of DC1 antigen lacks one of the tryptic cleavage sites; the DR heavy chain, the class I heavy chain, and beta 2 microglobulin lack all three proteolytic sites. Proteolysis of murine la antigens with chymotrypsin and trypsin generates similar fragments (I-E-like DR, I-A-like DC1) suggesting these cleavage sites are a general feature of class II antigen light chains.
Regulation of neutrophil inflammatory mediator release: chemotactic peptide activation of stimulus-dependent cytotoxicity.
Authors: English D , Lukens J N
Published: December 2022

Human neutrophils stimulated with phorbol myristate acetate (PMA) were able to damage human erythroleukemic K-562 target cells as assessed by a 3-hr 51Cr-release assay. Neutrophils from a patient with chronic granulomatous disease of childhood were ineffective in mediating PMA-stimulated cytolysis. Cytotoxicity was inhibited under anaerobic conditions as well as by catalase and several free radical scavengers. Superoxide dismutase, azide, and cyanide failed to inhibit PMA-dependent cytotoxicity. The influence of the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) on stimulus-dependent cytotoxicity was examined. Neutrophils preincubated with 1.0 X 10(-7) M FMLP did not display an increased cytotoxic response, but were markedly amplified in their ability to effect cytotoxicity with the addition of PMA. Enhancement of PMA-stimulated cytotoxicity resulted from a reversible cellular response to FMLP. The kinetics of the cytotoxic responses reflected the possibility that chemotactic peptide-treated neutrophils released cytotoxic mediators to a greater extent and at a faster rate than did untreated neutrophils. Although azide and superoxide dismutase did not inhibit cytotoxic responses of chemotactic peptide-activated neutrophils, the response was prevented by catalase and was markedly inhibited by several free radical scavengers. The ability of FMLP to enhance cytotoxic responses correlated well with its enhancement of PMA-stimulated chemiluminescence under a variety of conditions. In addition, the ability of PMA-stimulated neutrophils to mediate methane generation from dimethyl sulfoxide and ethylene generation from alpha keto-gamma-methiol-butyric acid (KMB)--assays that quantitate the generation of oxidizing radicals--was increased if the neutrophils were preincubated with FMLP. These results demonstrate that a chemotactic factor greatly potentiates the release of cytotoxic mediators from neutrophils upon stimulation with a nonchemotactic agent. The cytotoxic mediators appear to be products of oxidative metabolism. The cytotoxic potential of neutrophils that have responded to chemotactic stimuli to reach sites of inflammation may be activated in a similar manner.
C3 cleavage products stimulate release of prostaglandins by human mononuclear phagocytes in vitro.
Authors: Rutherford B , Schenkein H A
Published: December 2022

Human monocytes cultured for up to 48 hr in serum-free, chemically defined culture media released low levels of prostaglandin. C3b, C3bi, and C3c stimulated an indomethacin-sensitive, dose-responsive increase in the amount of monocyte prostaglandin released by 18 hr after treatment. Native C3 and C3d, which do not bind to monocyte receptors, failed to stimulate increased prostaglandin release. Lymphocytes, treated and untreated, produced 10(-2) to 10(-3) as much prostaglandin as the monocytes. These data support the concept that monocytes are a significant source of leukocyte prostaglandin. They also introduce an important new biologic function for the C3 fragments C3b, C3bi, and C3c.
Requirements for the opsonic activity of human IgG directed to type 6 group A streptococci: net basic charge and intact Fc region.
Authors: Fischetti V A
Published: December 2022

By two independent techniques for separating human opsonic IgG for group A type 6 streptococci into fast- and slow-migrating fractions, it was found that the opsonic activity was localized within the basic charge population. This charge dependence was found to be a characteristic of the IgG isolated from three individuals. When the fast- and slow-migrating IgG fractions were tested for their ability to bind to purified M6 protein, antibodies in both opsonic and nonopsonic populations exhibited binding activity, with the majority being located within the opsonic IgG in two of the three individuals; the third displayed greater binding in the nonopsonic population. The functional difference observed in the antibody populations to this M antigen may be a reflection of the net charge within the area of the antibody binding site, which suggests that the opsonic antibodies need to bind to acidic residues along the outer surface of the fibrillar M protein molecule. F(ab')2 fragments prepared from both human and rabbit type 6 opsonic IgG were still able to bind to the M6 molecule but were unable to mediate opsonization of type 6 streptococci. However, the F(ab')2 fragments had the capacity to enhance or amplify the opsonic activity of low concentrations of opsonic IgG molecules. The results suggest that the M protein molecule may function as an active inhibitor of phagocytosis and that F(ab')2 fragments from opsonic IgG have the capacity to neutralize the "active" determinants on the molecule, thus allowing lower concentrations of IgG with functional Fc receptors to mediate phagocytosis.