bluuebunny/crossref_metadata_2025 huggingface dataset:
The Identification of Actin as a Major Lymphocyte Component
Authors: Barber Brian H. , Delovitch Terry L.
Published: January 2023

Lymphocytes derived from pig mesenteric lymph node and mouse spleen have been shown to contain a major protein component which a) co-migrates on one-dimensional SDS-PAGE with purified pig muscle actin, and b) possesses the same isoelectric point as muscle action on two-dimensional gel electrophoresis. Further, limited proteolytic digestion of the lymphocyte actin-comigrating band by S. aureus V8-protease revealed extensive primary sequence homology with purified muscle actin. Based on the virtual identity of molecular size, charge, and primary sequence, we conclude that the actin-comigrating band represents lymphocyte actin. The appearance of lymphocyte actin as a nonspecific “contaminant” of immune precipitates has also been discussed.
Activation of the Classical Complement Pathway by Nephritic Factor Bound to the Alternative Pathway C3/C5 Convertase
Authors: Sobel Alain T. , Cooper Neil R. , Schreiber Robert D.
Published: January 2023

Nephritic Factor (NF), the potent alternative pathway activator, which is occasionally found in association with certain types of nephritis has recently been identified as an IgG class autoantibody specific for the C3 convertase of the alternative pathway. In these studies we have examined the possibility that the cell-bound NF-stabilized C3 convertase binds and activates the first component of the classical pathway of complement. bound C1q, and the extent of binding was dependent upon the number of NF molecules bound per cell and decreased parallel to the dissociation and release of NF from the cells. Interaction of C1 with bound NF resulted in its activation as shown by the proteolytic conversion of proenzyme C1s to its activated form C1s̅. As was the case with C1q binding, C1 activation was dependent on the number of NF molecules bound per cell. Thus the NF-stabilized C3 convertase binds and activates C1.
Comparisons of Myeloma Proteins from NZB and BALB/c Mice: Structural and Functional Differences of Heavy Chains
Authors: Loh E. , Hood J. M. , Riblet R. , Weigert M. , Hood L.
Published: January 2023

The N-terminal sequences from heavy variable regions of 47 myeloma proteins of the NZB mouse have been analyzed. Sixteen of these VH regions have unblocked α amino groups and have been analyzed over their N-terminal 20 residues by automatic sequence analysis. These sequence data along with the antigen-binding profiles and immunoglobulin class distribution are compared with comparable data from BALB/c myeloma proteins. These comparisons suggest that the NZB and BALB/c populations of myeloma proteins are distinct from one another. The genetic implications of this conclusion are discussed.
High Responder Rabbits to SRBC, a Familial Incidence
Authors: Bratcher Ray L.
Published: January 2023

Three litters of rabbits from our closed colony when immunized with multiple i.v. injections of SRBC gave unusually high titers compared to other rabbits immunized in a similar fashion. Data suggest that this response is a familial occurrence. The antisera from the high responding rabbits also exhibited other unique properties in addition to the high agglutinin titers. The early bleedings had significantly higher amounts of IgG anti-SRBC antibodies but similar amounts of IgM anti-SRBC antibodies relative to normal rabbits immunized with SRBC. The IgG antibodies from the high responders were low avidity antibodies and may exhibit limited heterogeneity.
Differences in the Behavior of the Membrane and Membrane-Associated Filamentous Structures in Normal and Chronic Lymphocytic Leukemia (CLL) Lymphocytes
Authors: Zucker-Franklin Dorothea , Liebes Leonard F. , Silber Robert
Published: January 2023

Ultrastructural studies of the membrane and membrane-associated cytoskeletal structures were carried out to help elucidate functional differences between normal and chronic lymphocytic leukemia (CLL) lymphocytes. A membrane-cross-linking ligand consisting of latex particles coated with anti-lymphocyte Ig permitted, because of its size, ultrastructural analysis of the site where the ligand had attached. In normal lymphocytes capped regions showed deep cytoplasmic invaginations and extensive polymerization of actin-like (7 nm) filaments in the adjacent submembranous zone. The presence of numerous bundles of intermediate filaments (10 nm) between the nucleus of the cell and the pole that carried the cap suggests a new role for these filaments in the capping process. CLL lymphocytes on the other hand showed deep cytoplasmic invaginations at sites of ligand binding over the entire cell surface. Actin-like filaments had also polymerized in the submembranous cytoplasm and had become attached to the plasma membrane by periodic cross bridges adjacent to these invaginations. However, organized arrays of intermediate (10 nm) filaments were not seen in CLL cells. Freeze-fracture studies after exposure of unfixed cells to glycerol revealed that the intramembranous particles (IMP) of normal lymphocytes had formed tight clusters whereas the IMP of CLL lymphocytes had either not been displaced at all or had formed only loose aggregates. This suggests the existence of a structural abnormality in the CLL Cell membrane that could interfere with IMP movement.
Helper Cells Activated by Allogeneic H-2K or H-2D Differences Have a Ly Phenotype Distinct from Those Responsive to I Differences
Authors: Swain Susan L. , Panfili Peter R.
Published: January 2023

Helper activity for the primary in vitro response to sheep erythrocytes was induced by recognition of foreign major histocompatibility antigens. The Lyt phenotypes of helper activity induced by differences in the whole haplotype, K or D antigens, I region antigen, or by differences at the M1s locus were determined. All allohelper cells expressed Ly1. In a single spleen cells suspension helper activity generated in response to the whole haplotype, I region, or M1s antigens was derived from an Ly2-negative population, whereas helper activity generated to K or D alone was derived from a population of Ly2-positive cells. Mixtures of anti-Ly1 and anti-Ly2 treated populations were unable to generate helper activity in response to K or D differences. Such helper activity was therefore dependent on the presence of Ly12 cells. It was concluded that the Ly phenotype of the helper cells is not determined solely by the T cell function but is influenced by the region of the major histocompatibility complex that is recognized. Possible interpretations of these findings are discussed.
Terminal Deoxynucleotidyl Transferase (TdT) in Chick Embryo Lymphoid Tissues
Authors: Sugimoto Masanobu , Bollum F. J.
Published: January 2023

The development of terminal deoxynucleotidyl transferase (TdT) was studied in the thymus and other lymphoid organs of embryonic and post-hatch chickens with an immunofluorescent procedure based on anti-TdT IgG purified on an immunoabsorbent column. TdT-positive cells were virtually absent in the thymus on day 11 of embryonic development and first became detectable on day 12. These cells then increased linearly to attain a plateau level of approximately 70% at about day 19. TdT-positive cells displayed network staining in the nucleus and some cytoplasmic staining. Most TdT-positive thymocytes were small, compared to TdT-negative cells.

A few TdT-positive cells (fewer than 1/105) were observed in the bursa of Fabricius in 20-day embryonic and 3- and 8-day post-hatch chickens. Positive cells were extremely rare in both the spleen and bone marrow. The brain and liver were TdT-negative.

These observations suggest that TdT-positive cells differentiate in situ from TdT-negative lymphoid cells in the thymus of embryonic chickens. The possible relationship of TdT to immune function is supported by the correlation of the kinetics of TdT-positive cell development in the present experiment with earlier observations on the morphologic, serologic, and functional development of embryonic chicken thymocytes.
Generation of T Helper Cells in Vitro
Authors: McDougal J. S. , Shen F. W. , Elster P.
Published: January 2023

In the thymocyte-macrophage (T-Mø) culture system for the in vitro induction of murine T helper cells, cortisone-resistant, nylon wool-purified thymocytes (CRNPT) are cultured with macrophages from antigen-primed donors. The particulate antigens, sheep erythrocytes (SRBC) or vaccine prepared from cell walls of Group A Streptococcus (SAV), were used to prime T-Mø cultures. The native CRNPT population contained Ly1, Ly23, and Ly123 cells in the proportions 45:3:46, but after 4 days of culture with antigen-pulsed macrophages the proportions were 88:2:2. The Ly1 T cell populations selected from such cultures by treatment with anti-Ly2 serum plus complement exhibited antigen-specific helper or suppressor functions in vitro, depending on the mode of the PFC assay: when assayed for their effect on T celldepleted spleen populations, helper function was manifest; when assayed with intact spleen cell populations, suppression was manifest. Our data do not indicate whether these two functions are mediated by the same Ly1 cells or by two subpopulations of Ly1 cells. The generation of Ly1 helper cells from CRNP thymocytes in 4-day T-Mø culture required the initial presence of the Ly123 members of this population. Manifestation of suppression by the same Ly1 population required the presence of Ly123 cells in the splenic populations used for PFC assay.
Cytotoxic Cells Induced During Lymphocytic Choriomeningitis Virus Infection of Mice
Authors: Welsh Raymond M. , Zinkernagel Rolf M. , Hallenbeck Linda A.
Published: January 2023

The target specificities of natural killer (NK) cells induced in mice by lymphocytic choriomeningitis virus (LCMV) were examined. All continuous and early passage cell lines tested were lysed by LCMV-induced NK cells, regardless of the target cell's origin (syngeneic, allogeneic, xenogeneic) or morphology (epithelia, fibroblast, lymphoblast, neuroblast, teratoma). Continuous cell line targets were lysed by LCMV- or vaccinia virus-induced NK cells from all tested strains of mice. Some target cells were more sensitive to lysis than other targets, but this relative sensitivity did not vary with the strain of mouse donating the effectors or with the time after inducing NK cells by the virus infection. LCMV-induced NK cells did not appear to recognize LCMV antigens on the surface of infected cells, nor could any correlation be made between retrovirus antigen expression and susceptibility to lysis. In contrast to this apparent lack of specificity in lysing continuous cell lines, we confirmed that some specificity could be demonstrated by using cold target inhibition experiments.

When primary peritoneal cells were used as targets, LCMV-induced NK cells from different strains of mice had restricted ranges of target specificities. NK cells from most homozygous mice lysed heterologous cells at much higher efficiency than isologous cells, but susceptibility to lysis was not directly related to H-2. In some strains of mice both isologous and heterologous targets were lysed. Spleen cells from F1 hybrid mice of certain strains lysed target cells from either parental strain. NK cell-mediated lysis of primary peritoneal cell targets did not require exogenous serum sources and was not significantly influenced by the presence of unlabeled target cells with varied susceptibilities incorporated into cytotoxicity assays.

We hypothesize that there may be two types of recognition mechanisms of the NK cell population: one that is “nonspecific,” resulting in the lysis of continuous or transformed cells, and another that is “specific,” resulting in a more selective lysis of primary cells. The vague specificities sometimes demonstrated against continuous cell targets by cold target inhibition experiments could be explained if the continuous cell line targets also expressed the specific recognition sites found on primary cells.
Mouse C4 (Ss): Three-Step Purification of Its C4c Fragment and Production of a Monospecific Antiserum
Authors: Ferreira Arturo , Nussenzweig Victor
Published: January 2023

We report here that a large fragment of mouse C4 (C4c) can be easily purified from serum with good recoveries, and used to produce a monospecific antiserum. The method of isolation of C4c is based on the observation that C4 is easily activated and fragmented in mouse serum. The fragments bind to the C4-binding protein (C4-bp), a newly described macromolecular component of the complement system. The high m.w. complexes were precipitated by dialysis of the serum against 0.1 M acetate buffer, pH 5.0, and purified by passage of the dissolved precipitate through a Sephadex G-200 column. The complexes were dissociated at high salt concentration, and the C4 fragments were isolated by passage of the mixture through a second Sephadex G-200 column. One of the C4 fragments (C4c) was used to immunize rabbits, and a monospecific antiserum was obtained, which showed a reaction of identity between C4 and Slp. Therefore the C4c fragment of C4 and the Slp protein are structurally related.
Establishment and Characterization of BALB/c Lymphoma Lines with B Cell Properties
Authors: Kim K. Jin , Kanellopoulos-Langevin Colette , Merwin Ruth M. , Sachs David H. , Asofsky Richard
Published: January 2023

Five spontaneously derived BALB/c tumors and one 1-ethyl-1-nitrosourea induced BALB/c tumor (BALENLM 17), have been established in culture and characterized according to their surface markers. K46, X16C, L10A, and BALENLM 17 are IgM+, IgG-, IgA-, Ig+ (staining with polyvalent anti-Ig), Ia+, Fc receptor+ and complement receptor-. A20 is IgM-, IgG-, IgA-, Ig+, Ia+, Fc receptor+, and complement receptor-. M12 is IgM-, IgG-, IgA-, Ig±, Ia-, Fc receptor+ and complement receptor-. All these lines are Thy 1.2 antigen negative, do not phagocytize latex particles, and grow as stationary suspension cultures. Therefore these six BALB/c lymphoid lines are presumably of B cell origin at various stages of differentiation. All these lines except X16C are tumorigenic in mice within a short period of time and have relatively short generation times in the range of 18 to 26 hr. Karyotype studies show that L10A, K46, X16C, and BALENLM 17 are hyperdiploid to hypotetraploid. A20 and M12 have near diploid chromosome numbers.
Killing of Schistosomula of Schistosoma Mansoni Coated with Antibody and/or Complement by Human Leukocytes in Vitro: Requirement for Complement in Preferential Killing by Eosinophils
Authors: Anwar A. R. E. , Smithers S. R. , Kay A. B.
Published: January 2023

The capacity of human eosinophils, neutrophils, and mononuclear leukocytes to damage schistosomula coated with antibody and/or complement has been studied by using an in vitro assay in which helminth destruction was visualised directly. Schistosomula and leukocytes were incubated either with i) antibody (Ab) alone (as baboon anti-schistosomula serum, or serum from patients with schistosomiasis or its IgG fraction); ii) complement (C) alone (by incubating schistosomula with fresh serum that led to activation by the tegument of the alternate pathway of complement as shown by immunofluorescence with anti-C3, but not anti-C4, both with normal and C2-deficient serum); or iii) the combination of Ab and C.

The efficiency of killing of schistosomula by granulocytes (i.e., mixtures of eosinophils and neutrophils) was 31 ± 7% for Ab alone, 52 ± 9% for C alone, and 70 ± 8% for Ab + C; control preparations were approximately 14%. Equivalent numbers of mononuclear leukocytes gave 22 ± 3%, 31 ± 5%, and 37 ± 4% killing with Ab alone, C alone, or Ab + C, respectively.

Schistosomula killing was directly related to the concentration of antibody with Ab alone and to the dilution of complement with C alone and Ab + C. Damage by granulocytes or mononuclear leukocytes was dependent on the ratio of effector cells to schistosomula in the three experimental systems.

Granulocyte-dependent damage was enhanced by the sequential addition, to schistosomula sensitized with antibody, of purified components of the classical pathway of complement and was directly related to the input of C3. This observation, together with the direct visualisation of C3 by immunofluorescence, suggests the participation of the classical pathway of complement in the Ab + C system.
Immunogenicity of Liposomal Model Membranes Sensitized with Dinitrophenylated Phosphatidylethanolamine Derivatives Containing Different Length Spacers
Authors: Dancey George F. , Isakson Peter C. , Kinsky Stephen C.
Published: January 2023

The immunogenicity of liposomes sensitized with dinitrophenylated (DNP) phosphatidylethanolamine (PE) derivatives, differing in the length of the spacer linking DNP to PE, has been examined. The magnitude of the anti-DNP response of mice was influenced by the number of methylene and amino groups interposed between DNP and the amino function of PE; generally, derivatives containing very short or long spacers were less effective immunogens than those of intermediate length. The epitope density of the derivative in the lipid bilayers, but not the dose, also affected the relative immunogenicity of the different derivatives. Incorporation of lipid A, which has been previously demonstrated to enhance liposomal immunogenicity, increased the response to those derivatives that elicited antibodies in the absence of this polyclonal B cell mitogen; however, the relative immunogenicity of the derivatives was not significantly altered. A good correlation was observed between the ability to bind low affinity IgG and IgM antibodies in vitro (determined by complement-dependent glucose release from liposomes) and in vivo immunogenicity. These results suggest that the ability of sensitized liposomes to elicit a response requires the extension of the antigenic determinant away from the surface of the lipid bilayer.
Early Stages of Human Marrow Lymphocyte Differentiation: Induction in Vitro by Thymopoietin and Ubiquitin
Authors: Kagan Walt A. , Siegal Frederick P. , Gupta Sudhir , Goldstein Gideon , Good Robert A.
Published: January 2023

To study early stages of human lymphocyte differentiation, bone marrow cells were physically separated according to their density and size by gradient centrifugation and then velocity sedimentation. The isolated cell fractions were incubated with putative inducing agents and then assayed for their expression of an array of surface differentiation markers. The inducing agents used were two polypeptides, thymopoietin (Tp) and ubiquitin (Ub), and the cyclic nucleotide, dibutyril cyclic 3′5′ adenosine monophosphate (cAMP). Tp, Ub, and cAMP each induced the ability to form sheep erythrocyte rosettes by small lymphocytes, which may thus represent T cell precursors. Ub and Tp induced rosette formation with mouse erythrocytes on lymphocytes of more heterogenous size, which may be “early” B cell precursors. Ub alone could induce surface IgM expression on small lymphocytes, which might be “late” B cell precursors. Both Tp and Ub induced Fc receptors on small lymphocytes. Complement receptors could not be induced on marrow lymphocytes by Tp, Ub, or cAMP. A number of lymphocyte precursors can thus be identified by their physical characteristics and their ability to respond to particular soluble factors with the expression of new differentiation markers.
Characterization of Thymus-Derived Lymphocyte Subsets in Acute Epstein-Barr Virus-Induced Infectious Mononucleosis
Authors: Haynes Barton F. , Schooley Robert T. , Grouse Jan E. , Payling-Wright Charles R. , Dolin Raphael , Fauci Anthony S.
Published: January 2023

Changes in thymus-derived (T) lymphocyte subpopulation numbers were studied in patients with acute and convalescent Epstein-Barr virus (EBV)-induced infectious mononucleosis (IM). T cell subsets were characterized by the presence of Fc receptors for IgG (TG), for IgM (TM) or by the absence of either receptor (Tnon-M, non-G). We found that in acute IM, total numbers of T and B lymphocytes were elevated (p < 0.01). Of the T lymphocyte subsets, the total number of Tnon-M, non-G lymphocytes was increased six fold compared to normal subjects (p < 0.001) and included the majority of the atypical T lymphocytes. The number of total TG and TM lymphocytes was moderately increased (p < 0.05). In convalescent IM patients, the number of total T cells remained slightly elevated (p < 0.02) whereas proportions and absolute numbers of B lymphocytes and T cell subsets returned to near normal levels. Thus, acute Epstein-Barr virusinduced IM is associated with a T lymphocytosis which is composed predominantly of atypical T cells which lack detectable Fc receptors for IgG or IgM.
Comparison of the immunosuppressive effect of fractionated total lymphoid irradiation (TLI) vs conventional immunosuppression (CI) in renal cadaveric allotransplantation.
Authors: Waer M , Vanrenterghem Y , Ang K K , Van der Schueren E , Michielsen P , Vandeputte M
Published: January 2023

Beginning in November 1981, eight patients with end stage diabetic nephropathy underwent renal cadaveric transplantation after TLI. Transplantation was done between 2 to 11 days after the end of a fractionated TLI to a total dose of 20 to 30 Gy. During the same observation period, 60 nondiabetic patients with end stage renal disease of different origin also received a cadaveric kidney graft, with a conventional regimen of immunosuppression that consists of anti-lymphocyte-globulin, tapering high doses of prednisone, and azathioprine. In the TLI-treated group only a low maintenance dose of prednisone (15 mg) was given. Immunologic monitoring was performed after transplantation at regular intervals and was compared in both groups. Phytohemagglutinin (PHA)-, concanavalin A (con A)-, and pokeweed mitogen (PWM)-induced blastogenesis, as well as the mixed lymphocyte reaction (MLR) and the cell-mediated lympholysis (CML) decreased progressively during the first months after conventional immunosuppression to 50% of the pretransplantation level, and remained there for the first year after transplantation. These tests were much more impaired after TLI (less than 15% of pre TLI value) and again no recovery occurred during the first year. Natural killer (NK) cell activity progressively decreased from a mean value of 53% lysis before transplantation to 15% lysis at the end of the first year after transplantation in the conventionally treated patients. In TLI-treated patients, however, the NK activity, which declined during irradiation from 46% specific lysis to 12%, recovered rapidly after TLI to reach levels of 35 to 40% of specific lysis from the second month on after TLI. In both groups of patients the ratio of helper-inducer (TH) to suppressor-cytotoxic (TS) lymphocytes, as determined with monoclonal antibodies, progressively declined during the first 2 mo after transplantation to a low value of about 1.2. In TLI-treated patients however, this fall progressed further, so that very low levels (less than 0.6) were noticed from the third month on after TLI. The decline of the TH:TS ratio after TLI is due to an absolute increase in the number of suppressor cells. This is in contrast with the conventionally treated patients, where the low ratio is mainly provoked by a more important decrease of the helper cell population. These changes in the balance between TH and TS subpopulations are more frequently associated with positive functional suppressor cell assays in the TLI-treated patients.(ABSTRACT TRUNCATED AT 400 WORDS)
Beta-interferon inhibits cell infection by Trypanosoma cruzi.
Authors: Kierszenbaum F , Sonnenfeld G
Published: January 2023

Preparations containing alpha/beta-interferon produced by L-929 cells were found to inhibit the capacity of bloodstream forms of Trypanosoma cruzi to associate with and infect mouse peritoneal macrophages or rat heart myoblasts. Marked reductions in the number of parasites per cell as well as in the percentage of cells associated with the trypanosomes were systematically observed in cultures of these cells that contained interferon. The inhibitory effect was abrogated in the presence of specific antibodies against alpha/beta-interferon, and purified beta-interferon induced a similar inhibitory effect, indicating that the active principle in the preparation was indeed interferon. Pretreatment of the parasites with alpha/beta-interferon reduced their infectivity for untreated host cells, whereas pretreatment of either type of host cell had no consequence on the interaction. The effect of interferon on the trypanosomes was reversible; the extent of the inhibitory effect was significantly reduced after 20 min, and was undetectable after 60 min when macrophages were used as host cells. Longer periods of time were required for the inhibitory effect to begin to subside (60 min) and to become undetectable or insignificant (120 min) when rat heart myoblasts were used. The results of additional studies performed with purified preparations of alpha- or beta-interferon revealed that only the latter was inhibitory of cell-parasite association. Because interferon is known to be produced shortly after T. cruzi infection and its administration has been shown to have a marked protective effect against this infection, our results suggest that the latter may involve inhibition of cell infection by interferon.
Assessment of antigen-specific restriction sites on Ia molecules as defined by the bm12 mutation.
Authors: Kanamori S , Walsh W D , Hansen T H , Tse H Y
Published: January 2023

To further delineate the functional defects of bm12 mice in antigen presentation, we analyzed antigen-specific T lymphocyte clones derived from B6 or bm12 for their ability to recognize antigens presented on either B6 or bm12 APC. Both complex antigens, such as PPD and GAT, and restricted antigens, such as beef insulin, were used. Our results indicate that for complex antigens, more than 50% of the B6 T lymphocyte clones recognized antigens only if presented by B6 APC, whereas the rest could not discriminate B6 from bm12 APC. Similarly, bm12 T lymphocyte clones responding to complex antigens could be divided into two groups depending on the sources of the APC. We have also isolated B6 T lymphocyte clones specific for the more restricted antigen, beef insulin, to which bm12 failed to respond. All B6 T lymphocyte clones could be stimulated only with B6 APC and not with bm12 APC. These data are consistent with the notion that there are antigen-specific association sites on the Ia molecule, and that complex antigens have more than one such association site. Furthermore, these studies demonstrate that both the gain and loss determinants associated with the bm12 mutation are recognized by a significant number of bm12 and B6 antigen-specific T lymphocyte clones, respectively, thus defining the importance of this region of the A beta polypeptide chain in antigen presentation.
Phylogeny of lymphocyte heterogeneity: the cellular requirements for in vitro mitogenic responses of channel catfish leukocytes.
Authors: Sizemore R C , Miller N W , Cuchens M A , Lobb C J , Clem L W
Published: January 2023

Cell separation and enrichment techniques were employed to isolate three distinct leukocyte subpopulations present in channel catfish (Ictalurus punctatus) peripheral blood. Surface immunoglobulin-positive (sIg+) and sIg- lymphocytes were separated by an indirect "panning" technique employing monoclonal antibodies reactive with channel catfish Ig. A third cell population composed of macrophages was isolated by adherence to baby hamster kidney cell microexudate-coated surfaces. Functional features of these three subpopulations were assessed by in vitro mitogenic responses to lipopolysaccharide (LPS) and concanavalin A (Con A). The results that were obtained indicated that the sIg+ cells responded only to LPS stimulation regardless of the presence or absence of macrophages. The sIg- subpopulation, however, responded to neither LPS nor Con A unless macrophages were present, in which case responses were obtained to both mitogens. The accessory cell nature of the macrophages was shown by experiments utilizing fixed numbers of one cell type mixed with varying numbers of another cell type. Furthermore, the accessory cell function was abrogated by passage through Sephadex G-10 and preincubation with L-leucine methyl ester. These studies provide further evidence that teleosts not only contain B and T cells akin to those in mammalian systems, but contain accessory cells (macrophages) as well.
H-41, a new minor histocompatibility locus. I. Histogenetic analysis.
Authors: Juretić A , Vucak I , Malenica B , Nagy Z A , Klein J
Published: January 2023

The B10.STA12 mouse congenic line inherited from the wild mouse parent not only the H-2w13 haplotype but also an allele at a minor H locus, which we designate H-41. This allele (H-41a) differentiates the B10.STA12 line from B10.STA10 and B10.LIB55, which carry identical H-2w13 haplotypes but a different H-41 allele (the H-41b, also present in the background strain C57BL/10Sn). The B10.STA12 and B10.STA10 lines reject each other's skin grafts and generate cytolytic T lymphocytes (CTL) after in vivo immunization and in vitro restimulation with cells of the partner strain. The B10.STA12 anti-B10.STA10 CTL react with B10.STA10, B10.LIB55, and B10.STA39 target cells and with cells of F1 hybrids between the responder strain B10.STA12 and strains C57BL/6, C57BL/10, C57L, BALB/c, A, AKR, WB, DBA/1, and DBA/2 but fail to react with (C3H x B10.STA12) F1 and (CBA x B10.STA12) F1 cells. The B10.STA10 anti-B10.STA12 CTL react with B10.STA12, B10.P, and C3H.NB cells but fail to react to (B6 x B10.STA10) F1 target cells. The CTL reactivity in both combinations is Dp restricted. The B10.STA10 anti-B10.STA12 CTL exhibit, in addition, a cross-reactivity with B10.SAA48 cells that may be directed at one of the alloantigens controlled by the H-2 haplotype of this strain.
The response of gut-associated T lymphocytes to intestinal viral immunization.
Authors: Issekutz T B
Published: January 2023

The cell-mediated immune response by the gut-associated lymphoid tissues to antigens within the intestinal tract is poorly understood. Our objective was to investigate the antigen-specific T cell proliferative response and cytotoxic T lymphocyte (CTL) response of cells from the GALT after enteric immunization with vaccinia virus. Lymphocytes able to proliferate in the presence of vaccinia virus in vitro were found in large numbers in mesenteric lymph nodes (MLN) 6 days after the injection of vaccinia virus into the lumen of the small bowel. The MLN at this time also contained vaccinia-specific CTL, but unlike the proliferating cells, which were found for several weeks after immunization, the CTL were demonstrable in the MLN for only a few days. Peyer's patches were found to contain neither antigen-stimulated proliferating cells nor CTL. The viral-specific proliferating lymphocytes from the MLN 10 days after immunization were sIg-, monoclonal antibody W3/25+, MRC OX-8- large lymphoblasts. The vaccinia-specific CTL were also large lymphoblasts, but they belonged to the W3/25-, OX-8+ subset. Thus, a strong T helper and cytotoxic T lymphoblast response is generated within the MLN after viral challenge of the gut.
A human hybrid myeloma for production of human monoclonal antibodies.
Authors: Kozbor D , Tripputi P , Roder J C , Croce C M
Published: January 2023

We produced somatic cell hybrids between human myeloma cells and a lymphoblastoid cell line that is hypoxanthine phosphoribosyl transferase-deficient and ouabain-resistant. These hybrids were phenotypically similar to the human myeloma parental cells and grew as well as the human lymphoblastoid parental cells. After counterselection in 6-thioguanine, mutants that were 6-thioguanine-and ouabain-resistant were obtained, one of which was used as a fusion partner with lymphoblastoid B cells that produce anti-tetanus toxoid (TT) antibodies. These hybrids secreted human anti-TT monoclonal antibodies in much larger amounts than the parental lymphoblastoid cells, and were stable for a period of over 10 mo until the present time. Thus, by hybridizing plasmacytomas with lymphoblastoid cells, we constructed a fusion partner that secretes large amounts of immunoglobulin (Ig), grows at a fast rate, has a high fusion frequency, and supports the production of monoclonal antibodies over long periods of time. Moreover, anti-TT antibody-producing hybrids have been grown as solid tumors in irradiated BALB/c nude mice and then adopted to ascites growth, producing 1 to 8 mg of human immunoglobulin per 1 ml of ascites fluid.
Resistance to tolerance induction is not prerequisite to development of murine SLE.
Authors: Izui S , Masuda K
Published: January 2023

Four different SLE-prone mice, NZB, (NZB x NZW)F1, MRL/MpJ-lpr/lpr, and male BXSB/MpJ, resisted the induction of tolerance to human IgG (HGG) at 5 wk, but not before 3 wk of age. However, female F1 hybrids between NZW and BXSB mice were easily made tolerant to HGG, although they develop an acute form of typical SLE similar to that seen in (NZB x NZW)F1 hybrid females. In addition, C57BL/6 mice bearing a mutant gene, lpr (lymphoproliferation), which produced significant amounts of various autoantibodies characteristic of murine SLE, were still as susceptible to tolerance induction as control C57BL/6 mice. In contrast, both (NZW x BXSB)F1 and (C3H x BXSB)F1 hybrid males carrying the abnormal Y chromosome of BXSB mice failed to become tolerant to HGG, although the extent of resistance to tolerance induction was less significant in (C3H x BXSB)F1 males than in (NZW x BXSB)F1 males. Our results suggest that 1) the defect in tolerance induction to heterologous IgG such as HGG is not necessarily required for the development of an SLE-like syndrome in mice; 2) the induction or enhanced production of autoantibodies by the lpr gene is not related to this cellular abnormality; but 3) a Y chromosome-associated factor from BXSB mice plays a significant role in the abnormality to resist tolerance induction as well as the acceleration of SLE. Our observations are consistent with the hypothesis that SLE may be based on highly specific abnormalities of immune responses to particular autoantigens, but not on a generalized breakdown of a tolerance mechanism.
Ly-2+ T cell enlargement and null cell proliferation occur at the onset of splenomegaly and autoantibody production in New Zealand Black mice.
Authors: Manohar V , Brown E M , Leiserson W M , Edison L J , Chused T M
Published: January 2023

Splenic T and B lymphocyte and null cell populations were analyzed in NZB mice as autoimmune disease developed during the first year of life. In the B lymphocytes, a progressive shift occurred from the surface IgD bright subset to the surface IgM bright subset. There was a slight increase in the ratio of Ly-2- to Ly-2+ T cells. Splenomegaly was not detected until after 40 wk of age and was primarily due to an increase in the number of null cells. This change was accompanied by an increase in the size, as determined by narrow-angle forward light scatter, of the Ly-2+ but not the Ly-2- T cells, an elevation of IgG-containing plasma cells, and the appearance of anti-erythrocyte autoantibody. The splenic B cell subset distribution and the enlargement of the Ly-2+ T cells were reflected in the peripheral blood, whereas the T cell subset ratio was not. The B cell subset alteration did not correlate with any of the other changes observed. Statistical associations were found between the ratios of T cell subsets, the enlargement of the Ly-2+ T cells, and the increased number of null cells, suggesting a linkage among those late changes that immediately precede the development of overt autoimmune disease.
Immunologic differentiation between E. coli and CHO cell-derived recombinant and natural human beta-interferons.
Authors: Colby C B , Inoue M , Thompson M , Tan Y H
Published: January 2023

The products of the human IFN-beta gene expressed in E. coli, Chinese hamster ovary (CHO) cells, and human fibroblasts appear similar when purified on a monoclonal antibody column and analyzed by reverse-phase HPLC, indicating little difference in their hydrophobic nature. SDS-PAGE differentiates E. coli-rHuIFN-beta ser (Mr = 17,000) from CHO-rHuIFN-beta and HuIFN-beta (Mr = 23,000), with glycosylation accounting for 26% of the apparent m.w. of the latter two proteins. CHO-rHuIFN-beta is preferentially neutralized by mouse monoclonal and monospecific rabbit polyclonal anti-HuIFN-beta antibodies, whereas E. coli-rHuIFN-beta ser is preferentially neutralized by goat polyclonal anti-E. coli-rHuIFN-beta antibodies. Adsorption measurements by a sensitive radioimmunoassay indicate that the binding of the three proteins to anti-HuIFN-beta antibodies is similar. The results show that all three molecules can be differentiated by the heteroclitic cross-reactivities of anti-HuIFN-beta and anti-E. coli-rHuIFN-beta antibodies to the antigens.
Mechanism of action of Cyclosporin A: inhibition of lymphokine secretion studied with antigen-stimulated T cell hybridomas.
Authors: Kaufmann Y , Chang A E , Robb R J , Rosenberg S A
Published: January 2023

We have employed bifunctional T cell hybridomas, which can be stimulated to secrete lymphokine(s) and lyse specific target cells, to analyze the effect of Cyclosporin A (CsA) on T cell helper and effector functions. We report here the effects of CsA on antigen- and lectin-induced lymphokine secretion. We have found that a pharmacologic level of CsA (10 ng/ml) blocks antigen- and lectin-driven interleukin 2 (IL 2) secretion without affecting cell proliferation. In addition, one monoclonal hybridoma that is induced by concanavalin A to secrete colony stimulating factors (CSF) as well as IL 2 is concomitantly blocked by CsA for production of IL 2 and CSF. Because the hybridomas grow constitutively and are devoid of functional IL 2 receptors, they permit analysis of the kinetics of the inhibitory response. We have shown that CsA blocks not only stimulation of lymphokine secretion but also ongoing IL 2 production, probably by interfering with the effective interaction of receptor and antigen. Thus, blocking of IL 2 secretion from preactivated cells by CsA occurs by 1 to 2 hr, the time required to stop IL 2 production by removal of Ag/Lectin stimulator. The results are consistent with a mechanism of action of CsA on T cells that involves a direct interference of CsA with binding of Ag to Ag-receptor and results in blocking of induction and active secretion of multiple lymphokines.
Lysine residues, but not carbohydrates, are required for the regulatory function of H on the amplification C3 convertase of complement.
Authors: Jouvin M H , Kazatchkine M D , Cahour A , Bernard N
Published: January 2023

Lysine epsilon-amino groups of human factor H were selectively converted to guanidino groups by treatment with 0.1 M O-methylisourea at pH 10.4. Guanidination resulted in a dose-dependent decrease in the capacity of the regulatory protein to accelerate decay dissociation of P-stabilized amplification C3 convertase sites, to serve as a co-factor for cleavage of cell-bound C3b by I, and to compete for binding of 125I-untreated H to C3b. Modification of approximately 75% lysine epsilon-amino groups suppressed 97% of H functional activity. Biochemical analysis of native H demonstrated a total carbohydrate content of 18.5% (w/w) and the presence in the molecule of 11 biantennary oligosaccharidic chains of the N-acetyl-lactosaminic type. Total desialation of H by using Clostridium perfringens neuraminidase, and total deglycosylation of desialated H by using beta-endo-N-acetylglucosaminidase resulted in a 1.5- to 2-fold increase in H activity on a weight basis. Deglycosylation did not alter the capacity of H to discriminate between activating and nonactivating surfaces of the alternative pathway. Thus, lysine residues are important determinants of the binding capacity of H for cell-bound C3b, whereas the carbohydrate portion of the molecule is not required for the regulatory function of the protein on the amplification C3 convertase.
Characterization of the murine TH response to influenza virus hemagglutinin: evidence for three major specificities.
Authors: Hurwitz J L , Herber-Katz E , Hackett C J , Gerhard W
Published: January 2023

Several hemagglutinin (HA)-specific T cell hybridomas were generated from five PR8 influenza virus-primed BALB/c mice by fusion with the thymoma BW5147 and were tested for IL 2 (interleukin 2) production in response to a large panel of serologically distinct influenza viruses and HA-protein fragments. The hybridomas (provisionally categorized in the helper T cell lineage) could be placed into three major specificity groups based on their reactivity patterns toward 43 antibody-selected PR8 virus mutants that had been shown previously to differ from parental PR8 virus by defined (usually single) amino acid substitutions in the HA1 polypeptide. Three hybridomas with specificity 1 failed to respond to the PR8 mutant RV6 (Glu115----Lys), two with specificity 2 failed to respond to PR8 mutant DV4 (Ser136----Pro), and two with specificity 3 responded to all antibody-selected PR8 mutants without exception. The three reactivity patterns could be used along with further biochemical evidence to demonstrate the presence of three distinct T cell determinants on the HA1 polypeptide. Priming of mice with one of these T cell determinants in the form of a synthetic peptide induced a T cell subset able to proliferate in vitro in response to HA and intact PR8 virus. Further analysis of the T cell hybridomas for reaction with 17 natural influenza virus isolates of the H1 subtype revealed additional differences in fine nominal antigen specificity among these T cells. However, two pairs of T cells whose members were isolated from different mice exhibited indistinguishable fine nominal antigen specificities. The latter analysis showed that natural antigenic drift (i.e., accumulation of point mutations) in the HA occurring between the years 1934 and 1957 and again between 1977 and 1980 altered two T cell determinants but left one T cell determinant unchanged. The shifts from H1 to H2 or H3 subtypes resulted in the alteration of all three T cell determinants. The present findings suggest that 1) immunization with influenza virus induces HA-specific T cells that recognize determinants different from those seen by anti-HA antibodies, and 2) the HA-specific T cell repertoire of adult virus-primed BALB/c mice is more restricted than the B cell repertoire.
Circulating monoclonal IgM proteins in B cell chronic lymphocytic leukemia: their identification, characterization and relationship to membrane IgM.
Authors: Qian G X , Fu S M , Solanki D L , Rai K R
Published: January 2023

Accumulated evidence indicates that there is a circulating monoclonal Ig protein related to the leukemic cell-associated Ig in the majority of patients with B cell chronic lymphocytic leukemia (CLL) despite the failure to demonstrate such a protein by conventional serum electrophoresis. Methodology has been developed to reveal these hidden monoclonal bands and to show that they are related to the leukemia-associated membrane Ig (mIg). Of nine CLL cases with stainable mIgM and without discernable plasma Ig bands, marked hypogammaglobulinemia was evident in six. In the other three, a significant amount of protein was present in the gamma region. IgM was isolated from the plasma of these patients by affinity chromatography with Sepharose-4B, conjugated with affinity purified anti-human IgM antibodies. One to 3 mg were isolated from 20 to 40 ml of plasma. Agarose electrophoresis revealed a monoclonal Ig band in the isolated IgM in all cases. Eight of these IgM proteins were analyzed by high-pressure liquid chromatography. Five were found to be pentameric IgM. In the remaining three, various amounts of monomeric IgM were detected. Attempts to make anti-idiotypic antibodies to the isolated proteins have been successful. Thus far, a rabbit anti-idiotypic antiserum was obtained in one case and two mouse monoclonal anti-idiotypic antibodies in two additional cases. Immunofluorescence analysis revealed that plasma IgM and mIgM shared similar idiotypic determinants. One other monoclonal antibody was shown to be specific for a V region marker of a minor Ig population. These findings indicate that B leukemic lymphocytes do secrete a small amount of IgM and lend further support to the thesis that the maturation defect in CLL is incomplete. It is also feasible to isolate the secreted IgM and to produce anti-idiotypic antibodies to them. In view of the potential therapeutic effect of anti-idiotypic antibodies, this may offer an alternative and efficient approach to generate a large panel of anti-idiotypic antibodies for clinical trials. The possibility also exists that this approach is applicable to other B cell proliferative disorders such as the non-Hodgkin B cell lymphomas.
Transfer of antigen-presenting capacity to Ia-negative cells upon fusion with Ia-bearing liposomes.
Authors: Coeshott C , Grey H M
Published: January 2023

The role of Ia in T cell activation was investigated by incorporating affinity-purified I-Ad molecules into synthetic liposomal membranes and by using these as antigen-presenting units. IL 2 production by I-Ad-restricted, chicken ovalbumin-specific T cell hybridomas was measured in a system in which antigen processing by the presenter was not required. I-Ad-bearing liposomes were found to have no antigen-presenting capacity. It was shown, however, that antigen-presenting capacity could be conferred on Ia-negative cells by fusion of these cells with liposomes bearing I-Ad molecules, together with Sendai virus envelope glycoproteins, as fusogenic agents. Both Ia-negative B lymphoma cells and mouse L cells were capable of antigen presentation of predigested ovalbumin after fusion with vesicles formed from phosphatidylserine and phosphatidylethanolamine in a 1:1 w:w ratio. The cell surface expression of the transferred Ia remained stable for at least 7 hr. These results indicate that Ia is the only additional cell surface molecule required, at least by Ia-negative B cell lymphomas and L cells, to convert them into effective antigen-presenting cells. This system should be useful in future studies of the cellular requirements for antigen processing and presentation.
Regulation of clones responding to dextran B1355S. III. The thymic-independent and thymic-dependent antibody responses.
Authors: Ward R E , Kohler H
Published: January 2023

The primary antibody response of different mouse strains challenged with two antigenic forms of alpha (1-3) dextran, dextran B1355S and dextran-hemocyanin, was examined. Only BALB/c mice responded with both kappa and lambda antibodies. The kappa to lambda ratio was affected by factors such as the antigenic form of dextran, the time at which the serum was analyzed, and the priming regimen. Surprisingly, priming with hemocyanin in adjuvant increased the kappa portion of the response not only to dextran-hemocyanin but also to dextran B1355S. Other strains of mice responded with only lambda antibodies. These results extend our previous results on the analysis of dextran-specific B cell precursors.