bluuebunny/crossref_metadata_2025 huggingface dataset:
Presence of T Cell-Associated Surface Antigens on Murine NK Cells
Authors: Pollack Sylvia B. , Tam Milton R. , Nowinski Robert C. , Emmons Sandra L.
Published: December 2022

The expression of T cell-associated surface antigens on natural killer (NK) spleen cells of C57BL/6 mice was evaluated by cytotoxic depletion experiments with alloantisera prepared against the Thy 1, Ly 1, Ly 2, Ly 5, Ly 6, and NK 1 antigens. The NK activity of these nonimmunized spleen cells for YAC-1 leukemia cells was dramatically reduced by antisera to the Ly 5 and NK 1 antigens. Variable results were obtained with anti-Ly 6 sera—certain pools of this antiserum decreased the NK activity, whereas other pools showed only negligible effects. The NK activity of the same cell suspensions was not affected by antisera to the Thy 1, Ly 1, and Ly 2 antigens. In parallel tests the T cell-associated cell surface antigens of alloimmune T killer cells were similarly evaluated by cytotoxic depletion experiments. In this case, the activity of these cells was consistently diminished by antisera to the Thy 1, Ly 2, Ly 5, and Ly 6 antigens, but not by antisera to the Ly 1 and NK 1 antigens. On this basis it was concluded that the NK cells expressed a restricted subset of T cell-associated alloantigens and therefore may have been derived from the T cell lineage of lymphocytes.
Studies on group A streptococcal M-proteins: purification of type 5 M-protein and comparison of its amino terminal sequence with two immunologically unrelated M-protein molecules.
Authors: Manjula B N , Fischetti V A
Published: December 2022

M-protein was isolated from group A, type 5, streptococci by limited proteolysis with pepsin and purified by chromatography on DEAE-Sephadex followed by gel filtration. The protein thus purified (Pep M5) was homogeneous by SDS-polyacrylamide gradient gel electrophoresis (apparent m.w.: 19,000), retained the capacity to remove opsonic antibodies from type 5 antiserum, and was capable of eliciting opsonic antibodies in rabbits. Its amino acid composition was very similar to that reported for M-proteins from other streptococcal types. The sequence of the first 29 amino terminal residues of Pep M5 was determined and compared with the reported amino terminal sequences of two immunologically unrelated M-proteins, namely, Pep M6 and Pep M24. The results revealed that, although the amino terminal sequences fo these three proteins differed from each other, some amino acid residues appeared to be conserved, suggesting a certain degree of structural relatedness among these M molecules. The possibility that this feature forms the molecular basis for the common antiphagocytic behavior of immunologically unrelated M-proteins is discussed.
A PMN-derived factor that enhances DNA-synthesis in PHA or antigen-stimulated lymphocytes.
Authors: Yoshinaga M , Nishime K , Nakamura S , Goto F
Published: December 2022

The DNA synthesis potentiation of a mouse polymorphonuclear leukocyte (PMN) factor was studied by using phytohemagglutinin (PHA)-stimulated syngeneic thymocytes. The partially purified PMN factor was nonmitogenic and exhibited maximum potentiation of thymocytes when added within 3 hr of PHA stimulation. PHA-stimulated, but not nonstimulated thymocytes absorbed PMN potentiation factor. 125I-labeled factor binding to stimulated thymocytes was observed. Our results suggest that an acceptor site for PMN factor may be generated on the thymocyte surface only after PHA stimulation.
Ability of NZW but not NZB antigen-presenting cells to support T cell proliferative response to DNA methylated albumin.
Authors: Papoian R , Talal N
Published: December 2022

The genetic contribution of the parental NZB and NZW mouse to the antigen-induced, macrophage-dependent, lymph node T cell proliferative response to d-DNA-mBSA was studied in F1 hybrid NZB/NZW (B/W) mice. Mice were immunized subcutaneously in the tail with denatured DNA-methylated BSA. Primed T cells from these mice were obtained after passage of immune periaortic and inguinal lymph node cells through nylon wool. The primed T cells were exposed in vitro to d-DNA-mBSA presented on spleen cells from BALB/c, DBA/2, NZB, NZW, or B/W mice. Immune B/W T cells gave a substantial proliferative response only in the presence of antigen-presenting cells from B/W or NZW mice. These results suggest that the response of B/W mice to DNA occurs primarily through the NZW H-2 haplotype of their own antigen-presenting cells.
Analysis and enrichment of murine natural killer cells with the fluorescence-activated cell sorter.
Authors: Tam M R , Emmons S L , Pollack S B
Published: December 2022

The antiserum anti-NK 1.1 defines an alloantigen specific for natural killer (NK) cells on normal C57BL/6 spleen cells (SC). With complement this antiserum lysed an insignificant percentage of SC, yet could deplete SC suspensions of NK effector cells. The antiserum was also used in this study to indirectly fluorescein label NK cells. The anti-NK 1.1 serum labeled 10 to 15% of nonadherent, nylon column passed SC. The labeled cells were analyzed on the fluorescence-activated cell sorter (FACS), by using flow fluorometry, and were found to be small to medium-sized cells. SC were sorted on the FACS into labeled (NK-1+) and unlabeled (NK-1-) populations, and assayed for NK activity on YAC-1 cells. When compared with control SC, labeled NK-1+ cells were enriched 4- to 13-fold in lytic activity, whereas unlabeled NK-1- cells had little if any NK effector function. Thirty to 60% of the labeled SC adhered to YAC-1 tumor cells in a visual target binding cell assay. The percentage of lymphocytes in the sorted NK-1+ population that bound YAC-1 cells was over 3-fold greater than in unsorted control SC preparations. NK-1- sorted cells did not bind YAC-1 targets. In a preliminary experiment NK-1+ sorted SC inhibited outgrowth of YAC in A/J mice, whereas NK-1- sorted cells did not.
Virus-induced immune complex disease: specific anti-viral antibody and C1q binding material in the circulation during persistent lymphocytic choriomeningitis virus infection.
Authors: Oldstone M B , Buchmeier M J , Doyle M V , Tishon A
Published: December 2022

Mice of several strains persistently infected with lymphocytic choriomeningitis virus (LCMV) mount continuous anti-LCMV immune responses leading to the formation and tissue deposition of immune complexes. Such mice carry infectious virus-immunoglobulin (presumably anti-LCMV antibody) complexes in the circulation. We have now determined that anti-LCMV antibody both complexed and free is found in the circulation of mice persistently infected with LCMV. This antibody reacts specifically against the three main LCMV structural polypeptides: nucleoprotein, 63,000 m.w. and two glycopeptides, GP-1 and GP-2 with m.w. of 45,000 and 35,000, respectively. A C1q binding assay was developed and found to be effective in measuring C1Q binding substances (presumably virus-anti-viral Ig complexes) in the circulations of several strains of mice persistently infected with LCMV. With different strains of mice, the levels, time of formation, and fate of C1q binding materials varied markedly. Formation of antibodies to LCMV was correlated with the detection of C1q binding materials. Mice (SWR/J) persistently infected with lactic dehydrogenase virus also form infectious virus-Ig in their sera but deposit minimal amounts of complexes in their tissues. In such mice, C1q binding substances did not form in the circulation.
Cellular requirements for generation of thymocyte stimulatory factor and characterization of its target cell.
Authors: Thoman M L , Weigle W O
Published: December 2022

The cellular requirements for the generation of thymocyte stimulatory factor, TSF, were defined, and the target cell for its activity was in part characterized. TSF production, which is triggered by concanavalin A stimulation of murine spleen cells, requires both an Lyt 1+23- cell and an Ia+ non-T accessory cell. The Ia+ cell does not bear surface Ig, is not killed by treatment with antithymocyte sera and C, and is partially depleted by Sephaadex G-10 filtration. TSF-augmentation of the thymocyte mitogenic response to Con A requires Lyt 1+, Ia- thymocytes, which are present in both the cortisone-sensitive and resistant populations. The relationship between TSF and other activities produced by Con A activation of spleen cells is discussed.
The use of cellular immunoadsorbents to prepare antibody that distinguishes between syngeneic surface antigens on two guinea pig hepatocarcinomas.
Authors: Mathews H L , Brunda M J , Minden P
Published: December 2022

Xenogenic antibodies with reactivity for surface determinants of the guinea pig line-10 hepatocarcinoma were isolated by using cellular immunoadsorbents prepared by coupling formalin-treated line-10 cells to diethylaminoethyl cellulose. Antibodies prepared in this manner exhibited a high degree of reactivity for line-10 surface determinants. These antibodies also reacted with surface determinants of the syngeneic line-1 hepatocarcinoma. Further specificity of antibody reactivity to the line-10 hepatocarcinoma was accomplished by passage of the antibodies through cellular immunoadsorbents prepared with syngeneic line-1 hepatocarcinoma cells. By direct binding studies, these antibodies showed significantly reduced reactivity for line-1 cells and no reactivity for guinea pig spleen cells or for the unrelated murine EL-4 lymphoma. By a competitive radioimmunoassay, these antibodies reacted only with determinants expressed on the surfaces of line-10 cells and not on syngeneic line-1, L2C, spleen, thymus; or xenogeneic EL-4 cell surfaces. In a similar manner, line-10-associated antigens were detected in ascites fluid derived from line-10 tumor-bearing animals. The sequential use of immunoadsorbents made of antigenically distinct but syngeneic tumor cells made it possible to prepare antibodies with restricted reactivity for line-10-associated antigens and should be applicable to the isolation of "tumor-specific" antibodies in other systems.
Structure and function of immunoglobulin domains. VIII. An analysis of the structural requirements in human IgG1 for binding to the Fc receptor of human monocytes.
Authors: Barnett Foster D E , Dorrington K J , Painter R H
Published: December 2022

A human mononuclear cell population, enriched with monocytes by adhering them to microexudate on Petri dishes previously used for fibroblast cultures was used in a homologous human system to form EA rosettes. IgG1, Fc, and subfragments of Fc representing the C gamma 2 and C gamma 3 domains were tested for their ability to inhibit rosette formation. Although Fc and IgG were equally effective in inhibiting rosette formation over the concentration range 10(-6) to 10(-5) M, all subfragments were inactive at these concentrations. At 10-fold higher concentrations the C gamma 2 fragment was still inactive; however, both the C gamma 3 fragments, pFc' and at C gamma 3, did show significant activity at this higher level. Reduction and alkylation diminished the inhibitory capacity of IgG 10-fold but had a lesser effect on Fc. In a parallel series of experiments the ability of IgG and Fc to inhibit granulocyte rosettes was found to be markedly diminished by reduction and alkylation in both cases. These experiments reveal differences between Fc receptors in different cells, confirm a role for the C gamma 3 homology region in monocyte Fc receptor recognition, but do suggest a requirement, either direct or indirect, for the C gamma 2 domain.
Evidence for a common mucosal immunologic system. II. Influence of the estrous cycle on B immunoblast migration into genital and intestinal tissues.
Authors: McDermott M R , Clark D A , Bienenstock J
Published: December 2022

The influence of the estrous cycle on the localization of plasma cell precursors in the genital mucosa was studied in CBA/J female mice by using an adoptive transfer system. Within 24 hr after transfer, [3H]-thymidine-labeled donor mesenteric lymph node (MLN) cells were observed in maximum number in proestral and estral cervices and vaginae (14.4 and 9.7 labeled cells/100 mm2 of tissue, respectively), and approximately 60% of these cells contained IgA and 25% IgG. During metaestrus and diestrus, however, few labeled MLN cells (4.0/100 mm2 of tissue) localized in the genital tract and this reduction affected the IgA-containing plasmacyte population in particular. In contrast, no change was noted in the frequency of labeled cells containing IgA in the small intestine over the course of the estrous cycle, although small but significant changes were noted in the frequency of labeled cells producing IgG. These results indicate that the hormonal changes during the estrous cycle may affect the localization of IgA plasma cell precursors in the genital tract and thus may alter the local mucosal immune response in this site.
Biosynthesis of lymphocyte surface IgD in the mouse.
Authors: Goding J W , Herzenberg L A
Published: December 2022

The synthesis of IgD was studied in mouse spleen cells by using [35S]methionine labeling followed by immunoprecipitation with monoclonal antibody and two-dimensional gel electrophoresis. After a 15-min pulse of [35S]methionine, a relatively basic form (IgD1) of apparent m.w. 59,000 was precipitated. Conversion into more acidic forms of m.w. 63 to 72,000 (IgD2) took place during a chase period of several hours. The acidic form was identical in mobility to that of IgD labeled by surface radioiodination, and was almost completely removed by treatment of intact cells with pronase. Neuraminidase treatment of the surface form (IgD2) produced a form resembling IgD1 in charge, but with no detectable change in m.w. Treatment of IgD1 with endoglycosidase H resulted in a form with an apparent m.w. of 50,000, whereas IgD2 was resistant to this enzyme. Both IgD1 and IgD2 bound to lentil lectin, whereas only IgD2 bound to Ricinus communis hemagglutinin, which binds to terminal galactose residues. These results indicate that IgD is synthesized as an incompletely glycosylated precursor possessing "high mannose" type oligosaccharide moieties, and passes relatively slowly through the cell. Shortly before surface appearance, galactose and sialic acid are added. No specific association with any other labeled protein was observed, and any IgD secretion was below the limits of detectability.
Cytotoxicity of anti-beta 2-microglobulin xenoantisera to human and murine myeloid progenitor cells and lack of cross-reactivity with colony-stimulating activity.
Authors: Fitchen J H , Ferrone S , Lusis A J , Cline M J
Published: December 2022

A previous report has suggested an antigenic relationship between beta 2-microglobulin (beta 2 mu) and granulocyte colony-stimulating activity (CSA). Since human myeloid progenitor cells (CFU-C) express HLA antigens and beta 2 mu is a known molecular component of HLA antigens, we wondered whether the reported effect of anti-beta 2 mu heteroantisera on in vitro granulopoiesis might result from cytotoxicity to CFU-C rather than from cross-reactivity with CSA. To test this, we used rabbit antibody reactive with human and murine beta 2 mu (anti-beta 2 mu). Treatment of human and murine bone marrow cells with anti-beta 2 mu and complement resulted in 95+% inhibition of CFU-C colony formation compared to controls. To test for an effect on CSA, anti-beta 2 mu was incubated with human and murine sources of CSA. After addition of goat anti-rabbit Ig antiserum to precipitate immune complexes and unbound anti-beta 2 mu, the supernatant fluid retained CSA but was no longer cytotoxic to CFU-C. These results indicate that human and murine CFU-C express membrane beta 2 mu and that anti-beta 2 mu antibody does not cross-react with human or murine CSA.
Collagen-induced polyarthritis in rats: a study of native type II collagen for adjuvant activity.
Authors: Cremer M A , Stuart J M , Townes A S , Kang A H
Published: December 2022

Collagen-induced polyarthritis in rats is a new experimental model that shares clinical and histologic features with adjuvant arthritis. To determine whether collagen-induced arthritis is a form of adjuvant disease and to further exclude contamination of collagen with an adjuvant substance, native type II collagen was studied for adjuvant properties. IgM and IgG PFC activity and PBMC [3H]TdR incorporation were studied in rats after injection with TNP-OA combined with IFA, IFA and CII, or CFA. In general, humoral and CMI responses to TNP-OA were lower in rats injected with IFA/CII compared with those with IFA; the presence of CII during primary immunization failed to significantly enhance PFC activity to TNP after a boost. CFA-injected rats gave maximal values in both studies. Mice pretreated with BII in the absence of oil gave PFC responses below control after sensitization with SRC. Furthermore, CII was unable to replace mycobacteria in the induction of EAE in rats and was devoid of mitogenic or polyclonal stimulatory properties. It is concluded that collagen-induced arthritis is a distinct entity from adjuvant arthritis and is dependent upon the unique immunogenicity of type II collagen in rats rather than upon an adjuvant effect.
Purification and biochemical characterization of an inhibitor of DNA synthesis produced by an Epstein-Barr virus-transformed B cell line.
Authors: Beneke J S , Qualtiere L F , Nesheim M C , Pearson G R
Published: December 2022

An inhibitory factor, which has been shown to suppress the uptake of 125I-iododeoxyuridine by both lymphoid and nonlymphoid cells, was isolated from the supernatant of an Epstein-Barr virus- (EBV) transformed B cell line (1605L) established from a cotton-topped marmoset. Purification of the inhibitor, which was produced in serum-free medium by crowded cultures of the 1605L cells, was achieved by DEAE-cellulose chromatography followed by preparative polyacrylamide gel electrophoresis. The apparent m.w. of the 1605L factor was determined to be 65,000 to 70,000 by SDS-polyacrylamide gel electrophoresis. The inhibitor was sensitive to digestion by trypsin and chymotrypsin but not RNase or DNase, indicating that it was protein in nature. Exposure of the 1605L factor to 56 degrees C for 1/2 hr or pH 2 for 48 hr at 4 degrees C destroyed its inhibitory activity. The biochemical characteristics and activity of the 1605L inhibitor distinguish it from Type I interferon and several other soluble immunologic mediators known to be produced by lymphoid cell lines.
Elastase-type proteases on the surface of human blood monocytes: possible role in amyloid formation.
Authors: Lavie G , Zucker-Franklin D , Franklin E C
Published: December 2022

A group of DFP-inhibitable serine proteases that are associated with the cell surface of human peripheral blood monocytes and degrade the amyloid precursor protein SAA has been partially characterized. Enzymes that resemble elastases in being inhibitable by Ac-Ala-Ala-Pro-Val-CH2Cl but differ from the secreted macrophage elastase in m.w. can be recovered from SDS gels in the region ranging from 58 to 72 x 10(3) daltons. These enzymes degrade SAA to a product similar in m.w. and antigenic properties to the amyloid A protein. When intact cells are labeled with 3H Ac-Ala-Ala-Pro-Val-CH2Cl at 4 degrees C for 3 min, a major 58 x 10(3) and two minor 48 and 65 x 10(3) dalton bands are seen. Another group of enzymes that digests SAA completely through a transient AA-like intermediate can be recovered from the 40 to 58 x 10(3) dalton region of the gels. These enzymes are only minimally inhibited by Ac-Ala-Ala-Pro-Val-CH2Cl. We suggest that both types of enzymes may be involved in the proteolytic processes that lead to amyloid formation in secondary amyloidosis.
Three M-components IgA lambda + IgG kappa n + IgG kappa h in one patient (DA): lack of shared idiotypic determinants between IgA and IgG, and the presence of an unusual kappa h chain of 30,000 M.W.
Authors: Bouvet J P , Liacopoulos P , Pillot J , Banda R , Tung E , Wang A C
Published: December 2022

Two apparently homogeneous electrophoretic bands were found in the serum of a patient (DA) with multiple myeloma. These M-components were identified as IgA-lambda and IgG-kappa paraproteins bearing different idiotypic determinants. Further analysis of the L chains showed that the lambda-chain was homogeneous but the kappa-chain could be separated by SDS-polyacrylamide gel electrophoresis into two different bands. Both of them were associated with gamma-chains but one (termed kappa n) had normal m.w. (24,500) whereas the other (termed kappa h) was larger (m.w. 30,000). Sugar content of the two DA IgG, as determined by anthrone reaction, was similar in DA IgG kappa n (0.73%) and in DA IgG kappa h (1.1%), clearly demonstrating that the difference in m.w. was not due to a large sugar chain. Furthermore, the peptide map of the kappa h chain included nine peptides absent in those of four other control kappa-chains. Sequence analysis showed that the first 25 N-terminal amino acids of the kappa n differed from those of the kappa h chain at positions 4, 5, 15, 18, and 21. Thus the two kappa-chains had different framework regions.
Two populations of Ia-like molecules on a human B cell line.
Authors: Lampson L A , Levy R
Published: December 2022

Evidence is presented for two populations of Ia-like molecules on the human B cell line Raji. Two monoclonal antibodies, L203 and L227, are described. Both recognize nonpolymorphic determinants on B cell antigens. Both immunoprecipitate 34,000- and 28,000-dalton material from 125I-labeled extracts of Raji, but L227 precipitates 25,000-dalton material as well. Immunodepletion experiments confirm that the antibodies react with two different populations of molecules.
The hepatitis B virus-associated delta antigen: isolation from liver, development of solid-phase radioimmunoassays for delta antigen and anti-delta and partial characterization of delta antigen.
Authors: Rizzetto M , Shih J W , Gerin J L
Published: December 2022

The delta antigen-antibody (delta Ag/anti-delta) system is associated with type B hepatitis in man. The delta Ag was extracted and solubilized by sonication and treatment with 6 M GuHCl from the nuclei of human liver obtained at autopsy of a delta-positive subject. A microtiter solid-phase radioimmunoassay was developed and used for the partial characterization of delta Ag. Equilibrium centrifugation, gel filtration and chemical and enzymatic treatments revealed delta Ag to be a protein of approximately 68,000 m.w. with a buoyant density of 1.28 g/cm3 in CsCl, stable to heat, acid, nucleases, and glycosidases, and inactivated by alkali and proteases. A standard preparation of delta Ag was used to develop a blocking solid-phase radioimmunoassay for anti-delta.
Interaction between the labile binding sites of the fourth (C4) and fifth (C5) human complement proteins and erythrocyte cell membranes.
Authors: Law S K , Lichtenberg N A , Holcombe F H , Levine R P
Published: December 2022

We have shown that the labile binding site of C3b interacts covalently with receptive surfaces. We report here an analogous study of the interaction between the labile binding sites of the closely related complement proteins, C4 and C5, with sheep erythrocyte membranes. We find that i) C4b binds covalently to cell surface components; ii) the bond between C4b and receptive molecules is hydroxylamine sensitive; iii) the alpha-polypeptide of C4b binds to receptive molecules; and iv) C5b does not interact covalently with cell surfaces.
Specificity of adoptive chemoimmunotherapy of established syngeneic tumors.
Authors: Cheever M A , Greenberg P D , Fefer A
Published: December 2022

To examine the specificity of adoptive chemoimmunotherapy (ACIT) of established syngeneic tumors, two noncross-reactive C57BL/6 tumors were studied: a Friend virus-induced tumor (FBL-3) and a chemically induced virus-negative tumor EL-4(G-). In vitro studies confirmed that these tumors are antigenically distinct by demonstrating that the cytotoxic responses of spleen cells from mice immunized in vivo and reexposed to tumor in vitro are immunologically specific. Studies of ACIT with cells from mice immunized in vivo demonstrated similar specificity. Mice receiving 5 x 10(6) FBL-3 on day 0 all died by day 13. Treatment on day 5 with cyclophosphamide (CY), 180 mg/kg, prolonged the median survival time (MST) to day 23. Treatment on day 5 with CY plus 2 x 10(7) normal nonimmune C57BL/6 cells or CY plus cells sensitized to EL-4(G-) had no additional effect on survival whereas 2 x 10(7) C57BL/6 cells sensitized to FBL-3 in vivo prolonged MST to day 64 and cured 13 of 32 mice. Similarly, mice given 2 x 10(5) EL-4(G-) on day 0 all died by day 16, and CY on day 5 prolonged the MST to day 22. As an adjunct to CY, 2 X 10(7) normal cells or cells sensitized to FBL-3 had a modest effect, prolonging the MST to days 37 and 36, respectively. However, treatment with CY plus 2 x 10(7) cells immune to EL-4(G-) cured 22 of 32 mice. The results demonstrate the immunologic specificity of ACIT of syngeneic tumors treated with immune syngeneic cells.
The activation of blast transformation and DNA synthesis in human peripheral blood lymphocytes by wheat germ agglutinin.
Authors: Gordon L K , Hamill B , Parker C W
Published: December 2022

Wheat germ agglutinin (WGA) has been shown to induce blast transformation and DNA synthesis in a minor subpopulation of human peripherl blood T lymphocytes (PBL). These cells enter DNA synthesis between 24 and 48 hr after stimulation, and demonstrate maximal blast transformation after 48 hr. In cultures of Ficoll/Hypaque purified PBL, the activation appears to be aborted 48 to 72 hr after WGA stimulation. After that time DNA synthesis is arrested and activated cells are rapidly lost from culture. The addition of N-acetyl glucosamine to cultures within 2 hr after WGA stimulation results in an increase in the proportion of cells activated and in the amount of DNA synthesized per cell. These experiments suggest that the abortive response may be due to the presentation of superimposed positive and negative signals.
A subpopulation of human polymorphonuclear neutrophils contains an active form of lactoferrin capable of binding to human monocytes and inhibiting production of granulocyte-macrophage colony stimulatory activities.
Authors: Broxmeyer H E , Ralph P , Bognacki J , Kincade P W , Desousa M
Published: December 2022

Subpopulations of human peripheral blood polymorphonuclear neutrophils (PMN) differing in functional capacity were separated by a rosetting procedure by using rabbit IgG antibody-coated sheep erythrocytes (EA). EA+ and EA- populations of PMN both contained lactoferrin (LF) as determined by radioimmunoassay. However, only LF (10(-16) to 10(-6) M) obtained from EA+ (= FC receptor (FcR)+) PMN was able to suppress the production of granulocyte-macrophage colony stimulatory activity derived from human monocytes. LF from EA- PMN was inactive at concentrations as high as 10(-5) M. Extracts of EA- PMN contained proteolytic enzymes, not apparent in FcR+ PMN, that inactivated the inhibitory activity of LF obtained from FcR+ PMN. These results were substantiated by immunofluorescence studies. LF from FcR+ PMN bound to monocytes whereas LF from EA- PMN demonstrated negligible binding to the monocytes and extracts from EA- PMN drastically decreased the capability of LF obtained from FcR+ PMN to bind to monocytes. These studies demonstrate functional heterogeneity of peripheral blood PMN and suggest a role for PMN subpopulations in the regulation of myelopoiesis.
Regulation by Fc fragments of the secretion of collagenase, PGE2, and lysozyme by mouse peritoneal macrophages.
Authors: Passwell J H , Dayer J M , Gass K , Edelson P J
Published: December 2022

Fc fragments of human IgG can stimulate resident mouse macrophages in culture to secret collagenase, to increase PGE2 secretion, and to decrease the secretion of lysozyme. Active synthesis and secretion were shown by the progressive accumulation of these products in the extracellular medium and inhibition of secretion by cycloheximide. A dose-dependent effect of Fc fragments was demonstrable. Brief exposure of cells to Fc fragments was sufficient to cause the macrophages to secrete collagenase and large amounts of PGE2 for prolonged periods of time, suggesting that a sustained activation rather than temporary modulation of the cells had occurred. Con A had similar effects on macrophage secretory activity. These findings indicate that proteins that bind to specific macrophage plasma membrane receptors may stimulate the secretion of products that promote the inflammatory response.
Effect of reduction on the structural, biologic, and immunologic properties of the third component of human complement.
Authors: Zemel E S , Cartwright-Harwick C M , Nilsson U R
Published: December 2022

A decrease of the hemolytic function of human C3 results from reduction with sodium borohydride (NaBH4) or with dithiothreitol (DTT) followed by alkylation with iodoacetamide. In the presence of 3.5 x 10(-2) M NaBH4, the loss of hemolytic activity (68%) is paralleled by a loss of C3b binding to EAC14oxy2 cells (67%), immune adherence (71%), and hemagglutinability (74%), but by no loss of C5 convertase function. Sulfhydryl analysis reveals that two disulfide bonds, both presumed to be situated in the C3 alpha subunit, are broken: one at 2.2 x 10(-2) M and one at 3.5 x 10(-2) M NaBH4. A contrasting inactivation profile is observed in the presence of 1.0 x 10(-1) M DTT. Loss of hemolytic function (76%) is paralleled by a decrease of C5 convertase function (87%), whereas C3b binding, immune adherence, and hemagglutinability are either unaffected or not significantly affected. Sulfhydryl analysis indicates the cleavage of four disulfide bonds; three bonds within C3 alpha and beta are broken at 5.0 x 10(-4) M DTT, and one additional bond within C3 alpha at 1.0 x 10(-3) M DTT. Antigenicity as determined by immunoelectrophoresis is unaltered at any of the above concentrations. The implication of these findings is discussed.
Effector cells in avian spontaneous and antibody-dependent cell-mediated cytotoxicity.
Authors: Fleischer B
Published: December 2022

Using effector cells from chickens and quails, antibody-dependent and spontaneous cell-mediated cytotoxicity (ADCC and SCMC) were studied in a 51chromium-release microcytotoxicity assay against erythrocytes and several tumor cells as targets. The effector cells mediating both ADCC and SCMC against tumor targets, as well as those active in ADCC against erythrocytes, exhibited similar characteristics: they were adherent cells of high density, which could be removed by treatment with carbonyl iron and magnet. The effector cell concentration was highest in the peripheral blood and lower in the spleen, whereas no activity was found in thymus and bursa. Lysis was detectable after 15 min of incubation, and the reaction was completed after a few hours. A rabbit antiserum was found to be able to induce ADCC against sheep erythrocytes, but it did so less effectively than a chicken antiserum. Studies on the target specificity of SCMC revealed a preferential destruction of xenogeneic lymphoma targets. These findings are discussed with respect to differences between the avian effector cells and the mammalian K and NK cells.
Regulation of thymus-independent responses: unresponsiveness to a second challenge of TNP-Ficoll is mediated by hapten-specific antibodies.
Authors: Brodeur P H , Wortis H H
Published: December 2022

CBA mice immunized with the TI-2 antigen TNP-Ficoll are unresponsive to a second challenge of the same antigen. In addition, spleen cells from unresponsive mice fail to respond to any dose of TNP-Ficoll in vitro. This hapten-specific unresponsive state is T cell independent and is due to hapten-specific IgM-mediated inhibition of B cell triggering. The mechanism of inhibition, although unknown, is unlikely to be antigen masking. In vitro responses to the TI-1 antigen TNP-LPS are not inhibited by concentrations of anti-TNP antibody, which completely suppress responses to TNP-Ficoll.
Antiidiotypic activity in the IgM fractions of mixed cryoglobulins.
Authors: Geltner D , Franklin E C , Frangione B
Published: December 2022

To search for human antiidiotypic antibodies, cryoglobulins from 11 patients with mixed cryoglobulinemia, 7 of whom had evidence of prior hepatitis B virus infection were separated on Sephadex G-200. Each of the IgM, 8 of which were IgM kappa, was tested with the F(ab')2 fragments prepared from each of the IgG fractions in a solid phase assay by using binding of 125I Staph A protein after the addition of rabbit anti-IgM. Unlike rabbit anti-Fab, which reacted approximately to the same extent with all F(ab')2 fragments, the IgM varied in their binding to F(ab')2 fragments, reacting with 2 to 10 antigens. Nine reacted with their autologous antigen and in 5 instances autologous reactivity exceeded that with heterologous F(ab')2 fragments. Reactivity was not related to prior exposure to HBV. Though absorption with Cohn fraction II F(ab')2 fragments generally abolished reactivity, 1 IgM protein continued to react with nine F(ab')2 fragments and 4 others with the F(ab')2 fragment from a single patient. Even when they contained only a single type of kappa-chain, the IgM appeared to contain multiple antibodies since absorption with solid phase Fc fragments or IgG removed anti-Fc rheumatoid factor activity but failed to affect binding to F(ab')2 fragments. Five of 7 IgM were able to bind the Fv region of a monoclonal IgM kappa-protein. Some of the anti-Fab antibodies were directed against idiotypes since addition of HbsAg to F(ab')2 fragments from HBsAb-positive IgG resulted in a marked decrease in binding of the IgM fraction in 6 out of 7 studies, whereas no decrease was noted in 6 experiments with F(ab')2 fragments that were HBsAb negative. The possible existence of antibodies against other idiotypes or other determinants in the Fab region cannot be excluded.
The expression of deoxyguanosine toxicity in T lymphocytes at different stages of maturation.
Authors: Cohen A , Lee J W , Dosch H M , Gelfand E W
Published: December 2022

Different human T cell populations were assayed for susceptibility of DNA synthesis to inhibition by deoxyguanosine. T lymphocytes from the thymus were most sensitive to inhibition of proliferation by deoxyguanosine (90% inhibition at 10 microM deoxyguanosine). This exquisite sensitivity of thymocytes appeared related to an enhanced ability of these cells for uptake and phosphorylation of deoxyguanosine to deoxyGTP and by their reduced ability to degrade accumulated deoxyGTP. Compared to more mature T lymphocytes and B cells, thymocytes contained the highest level of the salvage enzyme deoxynucleoside kinase and the lowest level of the nucleotide degrading enzyme, 5'-nucleotidase. The present study suggests that the levels of these 2 enzymes can serve as differentiation markers, identifying T cells at various stages of maturation, and that the loss of sensitivity to deoxyguanosine toxicity may be a stepwise process. Further, a deficiency in purine nucleoside phosphorylase may preferentially interfere with T cell maturation at an intrathymic stage of T cell differentiation.
Peanut agglutinin. VI. Identification of murine T lymphocyte subsets during ontogenesis by use of peanut agglutinin and Ly-6.2 antiserum.
Authors: London J
Published: December 2022

Fluoresceinated peanut agglutinin (PNA-FTC), Thy-1.2, and Ly-6.2 antisera were used to identify various murine T lymphocyte subpopulations during fetal and perinatal life. By means of double immunofluorescence labeling, we identified three T lymphocyte subsets: PNA+Ly-6.2+, PNA+Ly-6.2-, PNA-Ly-6.2+ having distinct distribution in the adult thymus and spleen. The study of the appearance of these three subsets in fetal liver, thymus, and spleen has shown that they did not occur similarly. The PNA+Ly-6.2+ subset was present in all three organs during fetal life and at a level rather similar to that of adult life; the PNA+Ly-6.2- subset was also present in the three fetal organs; conversely, the percentage of the PNA-Ly-6.2+ spleen cells was very low in fetal and neonatal life and increased significantly between 1 and 2 wk of age. Our results demonstrate that during fetal life, lymphocyte subsets are phenotypically differentiated as in adult life and that differentiation of T lymphocytes occurs preferentially through various lineages present during early ontogenesis. Moreover, quantitative modulation of some of these subsets is shown from fetal to adult life and may explain some of the immunologic immature features of young murine spleen.
Models for the production of stable hematopoietic chimerism across major histocompatibility barriers in adults.
Authors: Wegmann T G , Rosovsky J , Carlson G A , Diener E , Drell D W
Published: December 2022

Experiments presented in this paper indicate that DBA/2J (H-2d) mice parabiosed for more than 100 days to the F1 hybrid between DBA/2J and C3H/HeJ (H-2k) contain in their serum an anti-H-2k antibody that is preferentially reactive with hematopoietic tissue and is also capable of killing C3H/HeJ colony-forming stem cells in vivo (CFU-S). These findings make the antibody a likely participant in the "takeover" reaction, in which DBA/2J red and white cells eventually replace the F1 hybrid blood system. With these observations used as a basis, 10(8) DBA/2J spleen cells were injected along with anti-H-2Kk (anti-host) monoclonal antibody into 10-wk-old (DBA/2J x C3H/HeJ)F1 hybrid hosts. The recipients had a high surivival rate and exhibited long-term chimerism. Unlike the situation in the parabionts, the DBA/2J lymphoid cells in the injection chimeras were found to be unresponsive to C3H/HeJ alloantigens. This could result from the continued presence of F1 hybrid stimulator cells, which are missing in the parabionts. The injection chimeras indicate that one can establish stable chimerism in adult mice without irradiation or metabolic poisoning.
All classes of murine IgG antibody mediate macrophage phagocytosis and lysis of erythrocytes.
Authors: Ralph P , Nakoinz I , Diamond B , Yelton D
Published: December 2022

Antibody-dependent phagocytosis and lysis of sheep RBC by mouse peritoneal exudate cells were studied using IgG subclass fractions of sera prepared by protein A-Sepharose chromatography. Three IgG1, 3 IgG2a, 4 IgG2b, and 1 IgG3 hybridoma antibodies to RBC were also used. The results showed that some monoclonal IgG2a and IgG2b antibodies were more active than IgG1 or IgG3 preparations when normalized to hemagglutinating activity, but all IgG sources mediated both effector mechanisms. This is in accord with findings of strong Fc receptors on macrophages for the IgG2 isotypes and weaker binding by IgG1 and newly discovered IgG3 receptors. A monoclonal IgG2b with no hemagglutination titer showed normal cooperation with macrophages. In contrast, 2 hybridoma IgM were inactive in macrophage-mediated lysis and allowed only marginal ingestion. Similar results were obtained using several macrophage cell lines, mouse spleen and human peripheral blood mononuclear cells, showing that all these types of effector cells do not distinguish among (mouse) IgG subclasses in handling antibody-coated RBC.
Generation of cytotoxic lymphocytes to syngeneic tumor by using co-stimulator (Interleukin 2).
Authors: Mills G B , Paetkau V
Published: December 2022

Cytotoxic lymphocytes (CL) highly active against the syngeneic mastocytoma, P815, were generated from spleen cells of DBA mice cultured with co-stimulator (Interleukin 2) and P815. More CL activity was generated from spleen cells of P815 tumor-bearing mice than from spleen cells of normal mice. Thymus cells from tumor-bearing mice, however, did not produce increased CL activity. Most of the CL were Thy 1 and Ly 1 positive. The optimal culture conditions and kinetics were similar to those for the generation of allogeneic cytotoxic T lymphocytes. The cytotoxic activity against syngeneic P815 was similar in magnitude to the response of DBA spleen cells to allogeneic tumor lines and to the response of allogeneic CBA spleen cells to P815. Although CL generated from tumor-bearing mice did not lyse normal DBA cells, they did lyse, to a much lesser degree, a number of tumor cell lines other than the sensitizing P815. This nonspecific lysis was not H2 restricted nor was it restricted to tumors of lymphoid origin. Generation of nonspecific cytolytic activity was antigen independent, occurring in the presence of co-stimulator alone.
Generation of cytotoxic lymphocytes to syngeneic tumors by using co-stimulator (Interleukin 2): in vivo activity.
Authors: Mills G B , Carlson G , Paetkau V
Published: December 2022

Spleen cells from DBA/2J mice bearing the syngeneic tumor mastocytoma P815, incubated with co-stimulator (Interleukin 2) and P815 in vitro, were effective in killing P815 cells in vivo. Within 24 hr of injection, 1 X 10(7) cytotoxic lymphocytes (CL) killed most of 1 X 10(6) P815 cells. The host response to the tumor 7 to 9 days after the initial tumor injection was also greatly enhanced in mice that had received CL. This effect was potentiated in sublethally irradiated mice. CL were effective in mice with large tumors, overcoming suppressive factors that might be present. Under certain conditions, a significant fraction of CL-treated mice survived the P815 tumor indefinitely. These included i.p. CL given 2 days after a large dose (1 X 10(6)) of i.p. P815. In addition, some mice given i.v. and intra-tumor CL survived small doses (1 X 10(4)) of subcutaneous P815 cells. Long-term surviving mice remained resistant to challenge with 1 X 10(6) tumor cells (which is 10(4) times the normally lethal dose) for at lest 1 yr. No detrimental effects were noted even after injection of 5 X 10(7) CL per mouse.
Unique determinants associated with hybridoma proteins expressing a cross-reactive idiotype: frequency among individual immune sera.
Authors: Marshak-Rothstein A , Benedetto J D , Kirsch R L , Gefter M L
Published: December 2022

It is possible to prepare antisera that recognize individual members of a family of hybridoma proteins in which all members of the family express the cross-reactive idiotype (IdCR) characteristic of the A/J anti-Ars response. These hybridoma-specific reagents also recognize a small percentage of the IdCR+ anti-Ars antibodies, which constitute the normal A/J anti-Ars response. Thus, the previously reported sequence heterogeneity of our IdCR+ hybridoma proteins actually represents components of the normal A/J anti-Ars response. Determinants associated with one particular IdCR+ hybridoma protein, 36-71 (Id36-71), were found on approximately 0.01 to 5.0% of the IdCR+ antibodies in all of the A/J hyperimmune sera that were examined. Determinants associated with 2 other IdCR+ hybridoma proteins, 16-46-4-8 (Id16-46) and 44-10 (Id44-10) were also found in almost all A/J immune sera, again at various concentrations. These results indicate that the heritable trait (IdCR) is expressed as a large number of antibody sequences that probably arise from the somatic diversification of a limited number of germ line genes. The pattern of diversification is qualitatively similar among individual mice but varies with regard to the relative proportions of the individual antibody components.
Structural relation of murine "third locus" (H-2L) major histocompatibility antigens to the products of H-2K and H-2D loci.
Authors: Rose S M , Hansen T H , Cullen S E
Published: December 2022

The primary structures of the allelic products of H-2L, the murine "third locus" major histocompatibility antigen, were compared with the structures of the H-2K and H-2D antigens by tryptic-peptide analysis. The H-2Ld,H-2Lq, H-2Dd, H-2Dq and H-2Kd molecules were radiolabeled and isolated from spleen cell detergent lysates with specific alloantisera. Comparisons of the tryptic maps of the H-2L, H-2D, and H-2K molecules of the H-2d haplotype demonstrated that H-2Ld molecules were as different from H-2Dd molecules as H-2Dd molecules were from H-2Kd molecules. In contrast, the H-2Lq and H-2Dq molecules were found to be extremely similar and a second peptide separation method was required to discern structural differences between the 2 molecules. Comparisons of the allelic products H-2Ld and H-2Lq indicated possible structural differences despite the lack of a known serologic difference. These results, while raising questions about the genetic evolution of the H-2L locus relative to the H-2K and H-2D loci, demonstrate that the structure of the H-2L molecule is distinct from that of the H-2K and H-2D molecules. These structural analyses further substantiate the functional studies that define the H-2L molecule as the "third" major histocompatibility molecule of the mouse.
Characterization of multiple immunoglobulin mu-chains synthesized by two clones of a B cell lymphoma.
Authors: Sibley C H , Ewald S J , Kehry M R , Douglas R H , Raschke W C , Hood L E
Published: December 2022

We have identified 3 species of mu-chain synthesized by mouse WEHI 279 lymphoma cells that differ in cellular location, size and charge--mui (internal), mum (membrane), and mus (secreted). Lactoperoxidase and galactose oxidase labeling experiments localize the mum chain to the plasma membrane and the mui chain to the cytoplasm. Pulse-chase experiments demonstrate that the mui pool contains precursors to both mum and mus chains. Comparative peptide mapping studies and cell labeling in the presence of tunicamycin suggest that the mum chain is 2000 daltons larger than the mus chains and that the difference is due to polypeptide alterations rather than carbohydrate differences. The WEHI 279 lymphoma has differentiated while in culture to a more advanced stage in the pathway of B cell differentiation.
Hapten-specific human T cell lines. I. Functional analysis with respect to cytotoxic and helper activity.
Authors: Friedman S M , Hunter S B , Thomas Y , Chess L
Published: December 2022

Human PBL cells, sensitized in vitro to TNP or FITC-modified autologous stimulators, have been successfully grown in long-term culture by using TCGF derived from PHA-activated tonsil cells. Long-term cultures consist almost exclusively of T lymphocytes as defined by the formation of spontaneous rosettes with sheep erythrocytes and C-mediated lysis with heteroantibodies to human T cells (R alpha T H). The majority, but not all of these T cells, bear surface Ia antigens as detected by C-mediated lysis in the presence of alpha p23,30. Functionally, these cultures maintain high levels of hapten-specific cytolytic activity during many weeks of culture. In addition, a number of these T cell lines exhibit hapten specific proliferation when co-cultured with x-irradiated hapten modified autologous cells. During this proliferative response, helper factor(s) are released that trigger B cell differentiation into AFC. Data are presented that demonstrate that helper factor(s) production by T cell line cells is preferentially triggered by autologous, but nt allogeneic stimulator cells, bearing the relevant hapten.
Heterogeneity of the tolerant state in rats with long established skin grafts.
Authors: Smith R N , Howard J C
Published: December 2022

Tolerance in rats with long established allogeneic skin grafts brought about by the neonatal inoculation of hybrid lymphomyeloid cells is immunologically heterogeneous because 2 states of tolerance may be defined operationally as fully and partially tolerant. T cells from fully tolerant donors transfer tolerance and are unresponsive in MLC, but T cells from partially tolerant donors do not transfer tolerance and are modestly responsive in MLC. A state of tolerance may exist without detectable suppression. Therefore, tolerance, traditionally defined as the permanent acceptance of an allogeneic skin graft, does not reflect a uniform immunologic state.