bluuebunny/crossref_metadata_2025 huggingface dataset:
Size separation and polyclonal activation to immunoglobulin secretion of early precursors of B lymphocytes.
Authors: Deslauriers-Boisvert N , Mercier G , Lafleur L
Published: December 2022

In adult mammals the bone marrow is a major site of B lymphocyte production, which occurs through the differentiation of pluripotent stem cells. In vivo studies have permitted the description of intermediate stages of differentiation. Following the recent development of culture conditions and the discovery of many polyclonal activators, it is now possible to carry out in vitro a finer analysis of B lymphocyte precursors differentiation. Because in vivo studies have shown that precursors vary in size, adult murine bone marrow cells were separated according to size. Fractions were cultured with polyclonal mitogens and tested for their capacity to develop into Ig-secreting cells. With this assay system, up to six populations were detected in adult bone marrow that can differentiate along the B cell lineage. The characterization of those populations was made according to size, mitogen(s) responsiveness, and kinetics of maturation. Precursor frequencies estimated by limiting dilution analysis further suggest that in cultures of some fractions of the gradient, new lymphocytes are produced through differentiation of immature precursors, although in cultures of some other fractions B lymphocytes may arise by self-renewal of mature cells.
Thymic hormonal activity on human peripheral blood lymphocytes in vitro. II. Enhancement of the production of immune interferon by activated cells.
Authors: Shoham J , Eshel I , Aboud M , Salzberg S
Published: December 2022

The thymic extract TP-1 significantly enhanced the production of immune interferon (IF) by human peripheral blood mononuclear cells in Con A- or Raji- but not PHA-induced cultures. TP-1 effect was more pronounced under conditions of low IF production. The effect was evident before cell proliferation took place and without necessary concurrent effect of TP-1 on the blastogenic response. TP-1 had no detectable effect on nonactivated cultures. The enhancement of IF production by TP-1 to the degree demonstrated here necessitates consideration of its possible effects in every assay of thymic hormones that involves lectin or allogeneic stimulation of lymphocytes.
Molecular diversity of secretory IgA anti-DNP antibodies elicited in rat bile.
Authors: Montgomery P C , Lemaître-Coelho I M , Vaerman J P
Published: December 2022

Dinitrophenylated-type III pneumococcus has been used to induce biliary IgA anti-DNP antibody responses in rats. A solid phase radioimmunoassay was employed to quantitate the biliary IgA anti-DNP antibodies after subcutaneous, i.v., or intragastric immunization. All routes were shown to effectively elicit biliary IgA antibody responses. The microscale sucrose isoelectric focusing technique was employed to assess molecular diversity of the secretory IgA anti-DNP antibodies in bile. Subcutaneous and intragastric immunization resulted in complex antibody spectrotypes whereas i.v. immunization yielded restricted antibody spectrotypes, demonstrating that the route of immunization influenced the molecular diversity of the biliary secretory IgA antibodies. These findings are discussed in terms of mechanisms governing the induction of secretory antibodies.
In vitro immune response of human peripheral lymphocytes. V. PHA- and protein A-induced human B colony formation and analysis of the subpopulations of B cells.
Authors: Muraguchi A , Kishimoto T , Kuritani T , Watanabe T , Yamamura Y
Published: December 2022

Human B colony formation was observed with PHA or protein A as a mitogen. Preculture of B cells with mitogens in the presence of irradiated T cells for 3 days was a prerequisite for the induction of B cell colonies. About 300 to 700 colonies per 10(6) seeded cells were detected and a linear relationship between the number of cells seeded and the number of colonies developed was observed. Cells in PHA-induced colonies had surface IgM and/or IgD but no cytoplasmic Ig, whereas cells recovered from protein A-induced colonies had cytoplasmic Igs that were not only IgM but IgG and IgA. In each of the protein A-induced colonies, a sequential appearance of IgM-, IgG-, and IgA-producing cells was observed from day 3 to day 5, showing that IgM-, IgG-, and IgA-producing cells were derived from a single precursor cell. PHA and protein A had an additive effect on the number of colonies induced. About 50% of colonies induced in the presence of both PHA and protein A had Ig-producing cells. These results suggest that PHA and protein A may stimulate distinct subsets of B cells into colony formation.
Transfer of natural resistance to Marek's disease (JMV) with nonimmune spleen cells. II. Further characterization of protecting cell population.
Authors: Lam K M , Linna T J
Published: December 2022

Spleen cells from 8-week-old, nonimmunized donor chickens can transfer resistance to a supralethal dose of the JMV leukemia line of Marek's disease (MD) to newly hatched, highly susceptible, histocompatible recipients. The population of cells transferring resistance has previously been shown to be non-T, non-B, and nonmacrophage in nature. We present data here showing that heavily x-irradiated spleen cells were unable to protect recipients from leukemia challenge. Both complement receptor-bearing and -lacking cells could confer resistance to newly hatched recipients. Fc receptor-bearing cells conferred significant protection to recipients, whereas spleen cells depleted of Fc receptor-bearing cells were unable to protect chickens from death after JMV challenge. This indicates that the population of spleen cells, which is moderately radiosensitive and which possesses Fc receptors, is responsible for the transfer of natural resistance to the malignancy in vivo.
Polyclonal activation of human lymphocytes in vitro. I. Characterization of the lymphocyte response to a T cell-independent B cell mitogen.
Authors: Schuurman R K , Gelfand E W , Dosch H M
Published: December 2022

The staphylococcal cell wall component protein A (SpA) and formalinized, Cowan I strain Staphylococcal organisms (STA) were compared with the lectins phytohemagglutinin, concanavalin A, and pokeweed mitogen for their ability to trigger proliferation of normal human lymphocytes, lymphocyte subpopulations, and cells from patients with primary immune deficiency diseases. SpA was found to be a potent T cell mitogen, very similar to the other lectins tested. It failed to stimulate purified non-T cells and peripheral blood lymphocytes from patients with different forms of severe combined immunodeficiency disease (SCID). STA, treated to prevent the leakage of soluble SpA during culture, exclusively stimulated non-T cells: the responding cell population was characterized to be E-rosette negative but positive for C3 receptors, surface Ia, a receptor for STA itself, and likely carried surface immunoglobulin. Normal responses to STA were found in patients with the adenosine deaminase-positive form of SCID. In 18 patients with humoral immune deficiency syndromes, the presence of STA responses was correlated with the presence of circulating, surface immunoglobulin-bearing cells. A commercial STA preparation was rendered B cell specific after reformalinization, a procedure that eliminated the shedding of soluble SpA under culture conditions.
Effects of thymectomy or androgen administration upon the autoimmune disease of MRL/Mp-lpr/lpr mice.
Authors: Steinberg A D , Roths J B , Murphy E D , Steinberg R T , Raveche E S
Published: December 2022

MRL/lpr and NZB X NZW F1 mice were neonatally thymectomized or treated with androgens from 2 weeks of age, and their natural histories were studied. Neonatal thymectomy of MRL/lpr mice led to marked reduction in the usual massive lymphadenopathy as well as significant reduction in antibodies to native DNA and prolonged survival. In contrast, neonatal thymectomy of NZB X NZW F1 mice led to accelerated disease. Androgen therapy of both mice led to reduced anti-DNA and prolonged survival. This occurred in the MRL/lpr mice without significant reduction in lymphadenopathy. Taken together, these studies suggest that different T cell abnormalities may underlie the development of autoimmunity in MRL/lpr and NZB X NZW F1 mice, but that disease may be expressed through a common androgen-sensitive pathway.
The effect of anti-immunoglobulin antibodies on the in vitro proliferation and differentiation of normal and neoplastic murineB cells.
Authors: Isakson P C , Krolick K A , Uhr J W , Vitetta E S
Published: December 2022

In these studies, we have compared the effects of anti-mu and anti-delta antibodies on normal and neoplastic murine B cells. Spleen cells from normal BALB/c mice or from mice carrying the BCL1 tumor were treated with F(ab')2 fragments of affinity purified anti-mu or anti-delta antibodies and the effec of the antibodies on cell proliferation was assessed. F(ab)'2 fragments of both antibodies induced proliferation of normal adult B cells but inhibited (anti-mu) or had no effect (anti-delta) on proliferation of BCL1 cells. In addition, both antibodies further enhanced the LPS-induced proliferative response of cells from adult mice. In contrast, the LPS responses of neonatal cells and BCL1 cells were inhibited by F(ab')2 anti-mu antibodies. These data indicate that interaction of antibody with surface IgD as well as surface IgM can activate adult B cells. In addition, the results provide further evidence that BCL1 cells are neoplastic analogs of immature B cells since the tumor cells, like neonatal cells, are not activated after treatment with anti-Ig antibodies.
A predominant idiotype on anti-hen egg white lysozyme antibodies from diverse mouses strains.
Authors: Benjamin C D , Miller A , Sercarz E E , Harvey M A
Published: December 2022

A predominant idiotype was detected on almost all antibody molecules directed against the multideterminant antigen, hen egg white lysozyme (HEL). This idiotypic set was defined by rabbit and quinea pig antiidiotypic antisera raised against a rare cross-reactive antibody population isolated and from a heterogeneous BALB/c anti-human lysozyme (HUL) humoral response. This isiotype was fond on the majority of all HEL-reactive antibody from all strain tested; its presence was independent of Ig-1 allotype of H-2 haplotype. The idiotype was also present on the majority of anti-HEL antibody from nonresponder mice immunized with keyhole limpet hemocyanin coupled to HEL. The idiotype was not found on normal mouse immunoglobulin or on antibodies specific to two other proteins, HUL and ribonuclease, which are similar to HEl in size and charge. Individual idiotype-positive anti-HEL antibody populations exhibited specificity differences when analyzed for reactivity toward other avian lysozymes, suggesting that distinct clonal products shared the idiotype specificities. Fractionation of heterogeneous anti-HEL antibodies by preparative isoelectric focusing yielded distinct antibody fractions that were demonstrated to share the relevant idiotype determinants. The data are discussed in terms of the HEL peptide specificites of idiotype-positive and -negative antibodies.
The mouse-primed lymphocyte typing test. III. Dissociation of T lymphocyte-stimulating determinants and antibody-defined specificities of the I region-associated Ia antigens.
Authors: Peck A B , Klein J , Wigzell H
Published: December 2022

The mouse-primed lymphocyte typing (mPLT) assay, based on the sequential reactivation of specific immunocompetent alloantigen-reactive T blast cells in secondary mixed leukocyte culture (MLC) has been utilized to define the major histocompatibility complex (MHC) I region-, or Class II-associated lymphocyte-stimulating (LS) determinants. The test panel of secondary stimulating cells has been expanded to include 28 B10.W lines (mouse strains carrying MHC regions derived from wild mice); thus, nearly 40 "independently derived" MHC haplotypes are now represented. Data obtained by using mPLT cells generated in primary MLC between I-AB or I-EC subregion disparate strain combinations reveal that 1) an absolute correlation between expression of the serologically defined Ia specificities and capacity to induce subsequent secondary MLC does not exist; 2) based on the apparent genetic derivations of the I-AB and I-EC subregions, T lymphocytes most likely recognize either the beta-chain per se or an interaction product of the alpha- plus beta-chains; and 3) multiple restimulations of mPLT cells with cells expressing cross-reactive serologically defined Ia specificities fail to select for cells with increased reactivity against shared specificities. Based on these observations, we conclude that t and B lymphocytes recognize different antigenic moieties expressed on the MHC Class II antigens.
Relative susceptibilities of T cell subsets involved in delayed-type hypersensitivity to sheep red blood cells to the in vitro action of 4-hydroperoxycyclophosphamide.
Authors: Kaufmann S H , Hahn H , Diamantstein T
Published: December 2022

4-Hydroperoxycyclophosphamide, a derivative of cyclophosphamide, in aqueous solution yields 4-hydroxycyclophosphamide, which acts on lymphocytes in vitro. This compound was employed to determine the relative susceptibilities of T cell subsets involved in delayed-type hypersensitivity to sheep red blood cells in vitro. The following ranking order could be established: precursors of suppressor T cells > antigen-activated suppressor T cells > T cells mediating DTH. These data provide direct in vitro evidence that the enhancement of the cellular immune response to sheep red blood cells by cyclophosphamide is due to selective inactivation of suppressor T cells or their precursors, respectively.
Isolation and characterization of feline C3 and evidence for the immune complex pathogenesis of feline infectious peritonitis.
Authors: Jacobse-Geels H E , Daha M R , Horzinek M C
Published: December 2022

Infections of cats with feline peritonitis (FIP) virus are usually inapparent but may lead to fatal polyserositis. We have recently advanced the hypothesis that immune complexes play an essential role in the pathogenesis of the condition. To support this hypothesis, the role of the third component of complement in FIP was investigated. In the present paper, the isolation of C3 from normal cat serum and some of its physical and immunologic properties are described. The final protein had an apparent m.w. of 185,000 and was composed of 2 polypeptide chains with m.w. of 128,000 and 71,000, respectively. When tested against whole cat serum, an antiserum raised in rabbits against purified C3 recognized only 1 protein whose identity with C3 was established. With the aid of this antiserum, depositions of C3 in renal glomeruli of FIP-affected cats were demonstrated by immunofluorescence. Their localizaton coincided with that of deposited IgG, thereby supporting the concept of an immune complex pathogenesis of FIP.
Free sulfhydryl groups of rabbit secretory IgA.
Authors: Elliott B W , Friedenson B , Knight K L
Published: December 2022

The reaction of sIgA with 5,5'dithiobis (2-nitrobenzoic acid) or with 14C-iodoacetamide revealed an average of 1 mol -SH per mol sIgA under nondenaturing conditions, whereas an average of 7.5 mol -SH was found under denaturing conditions. These -SH groups were found in both the sIgA-f and sIgA-g subclasses; in the sIgA-g subclass, the free -SH groups were found in the Fab alpha portion but not in the Fc alpha portion of the molecule. To determine whether the noncovalently bound polypeptide chains in sIgA contained free -SH groups, sIgA was alkylated with 14C-iodoacetamide and subjected to gel filtration in 6 M guanidine-HCl. This treatment liberated several different polypeptides that were identified as secretory component (SC), L chain dimers, and L chain monomers; measurement of radioactivity incorporated in each of these noncovalently bound chains showed that SC and the L chain dimers did not contain free -SH groups but that an average of 1 -SH group per L chain monomer was found. Of the covalently bound polypeptide chains, J chain and SC did not have detectable -SH groups, whereas the L chains and alpha-chains had 0.3 and 1.2 mol -SH per mol protein, respectively. The identification of free -SH groups in the Fab portion of the sIgA-g subclass and their preferential localization within only some of the L chains reflect the structural heterogeneity of sIgA; these -SH groups presumably arise during biosynthesis.
Chemical enhancement of tumor immunogenicity.
Authors: Eggers A E , Hibbard C A , Civin C I , Wunderlich J R
Published: December 2022

C57BL/6 spleen cells immunized in vitro against syngeneic methylcholanthrene-induced sarcoma cells(MC-1 cells) modified with various chemical reagents show cytotoxic activity against unmodified MC-1 cells in a short-term 51Cr release assay, whereas unmodified MC-1 cells are nonimmunogenic. The effector cells cross-react widely with many other fibroblastic and epithelioid tumors, even those that are not H-2 matched, as well as with some nonneoplastic cells. Priming mice in vivo with a hapten leads to enhanced anti-tumor cytotoxicity developed by spleen cells from these mice immunized in vitro against tumor cells modified with the same hapten. Cytotoxic activity is largely, but not completely, removed by treatment with anti-theta serum and C. The existence of suppressor cells that can inhibit an anti-tumor response is demonstrasted in in vitro immunizations of mixtures of spleen cells from normal mice and mice primed with mitomyhcin-treated tumor cells, the latter suppressing the former.
Proteolysis of the C5b-7 complex: cleavage of the C5b and C6 subunits and its effect on the interaction of the complex with phospholipid bilayers.
Authors: Yamamoto K I
Published: December 2022

The present study was conducted to gain insight into the process of assembly of the C5b-7 complex on the phospholipid bilayer. The C5b-6 complex, C7, and 2 different forms of the C5b-7 complex (the C5b-7 complex either bound to artificial phospholipid bilayers or in fluid phase) were digested with trypsin, and the resulting products were analyzed by SDS-polyacrylamide gel electrophoresis. The alpha'-chains of C5b and C6 in the C5b-6 complex and C7 were susceptible to trypsin. The formation of the C5b-7 complex from the C5b-6 complex and C7 was accompanied by alteration in susceptibility to trypsin to C7: the C7 subunit of the fluid phase complex as well as of the complex bound to phospholipid vesicles is not susceptible to trypsin. On the other hand, the alpha'-chains of C5b and C6 in the C5b-7 complex bound to phospholipid vesicles as well as in fluid phase remained accessible and susceptible to trypsin, and cleaved as in the C5b-6 complex. Upon ultracentrifugation in a sucrose density gradient, the proteolyzed C5b-7 complex in fluid phase dissociated into subunits. However, none of the subunits in the proteolyzed C5b-7 complex bound to phospholipid vesicles dissociated from vesicles upon sucrose density gradient ultracentrifugation. Thus, the molecule of the C5b-7 complex became stable and resistant to proteolytic dissociation upon binding to the phospholipid bilayer. It is proposed that each subunit of the C5b-7 complex participates in the stable interaction of the complex with the phospholipid bilayer. In addition, the possible significance of the alteration in susceptibility to trypsin of C7 associated with the formation of the C5b-7 complex is discussed.
Lymphocyte recognition of lymph node high endothelium. I. Inhibition of in vitro binding by a component of thoracic duct lymph.
Authors: Chin Y H , Carey G D , Woodruff J J
Published: December 2022

Rat thoracic duct lymphocytes (TDL) adhere selectively to HEV when overlaid onto glutaraldehyde-fixed frozen sections of lymph node. The results presented demonstrate that thoracic duct lymph contains a factor capable of inhibiting this reaction. A crude preparation of inhibitory factor was isolated from cell and chylomicron-free lymph by (NH4)2SO4 precipitation (60 to 80% saturation). Sephacryl S-200 chromatography revealed 4 major peaks. The inhibitory factor was detected in peak I but not in the others. Similarly peak I but not peak II or III inhibited HEV binding of lymphocytes from lymph nodes and spleen. Peak I emerged near aldolase (m.w. 160,000) and was enriched approximately 50-fold compared to the crude preparation. In addition, experiments employing a rabbit peak I antiserum provided evidence that molecules responsible for inhibition were antigenically related to structures present on TDL but not on thymocytes. It is suggested that inhibitory factor possesses an affinity for lymphocyte binding sites of high endothelium and is derived from shed TDL surface components that play a role in adherence to HEV of lymph nodes.
Lymphocyte recognition of lymph node high endothelium. II. Characterization of an in vitro inhibitory factor isolated by antibody affinity chromatography.
Authors: Chin Y H , Carey G D , Woodruff J J
Published: December 2022

Thoracic duct lymph contains a factor that inhibits in vitro adhesion between lymphocytes and high endothelial cells. Crude inhibitory factor was isolated from lymph by (NH4)2SO4 precipitation and partially purified by using an immunoabsorbant column of rabbit anti peak I Ig coupled to Sepharose 4B. Antibody affinity chromatography separated the HEV binding inhibitory factor from the bulk of protein in the crude preparation; activity was enriched 50-fold. The results show that the effect of the factor was exerted on high endothelial cells; both glutaraldehyde-fixed HEV and unfixed mouse HEV were susceptible to the action of this material. In contrast, the HEV binding properties of lymphocytes were unaffected by the factor. Inhibitory activity was destroyed by treatment with trypsin or exposure to 100 degrees C but was unaffected by incubation at 56 or 70 degrees C for 30 min. In addition, the factor bound to lentil lectin and was eluted with alpha-methyl-D-mannoside. Together these findings indicate that the HEV binding inhibitory factor is a glycoprotein.
Use of benzoyl-L-phenylalanyl-L-valyl-L-arginine (3H) methyl ester as a sensitive and selective substrate for the human lymphokine, leukocyte migration inhibitory factor (LIF).
Authors: Bendtzen K , Rocklin R E
Published: December 2022

Human leukocyte migration inhibitory factor (LIF) appears to be a serine esterase and protease exhibiting specific affinity towards arginine esters and amides. On the basis of indirect evidence that an amide of the oligopeptide benzoyl-phenylalanyl-valyl-arginine might have high and selective affinity for LIF, we prepared an ester of this oligopeptide, benzoyl-phenylalanyl-valyl-arginine (3H) methyl ester (3H-BPVAME) for the direct measurement of LIF activity in a double-phase radio-enzyme assay. The hydrolysis of 3H-BPVAME followed enzyme-substrate kinetics in that the reaction was time-, temperature-, pH-, and concentration-dependent. 3H-BPVAME rpoved to be more selective and approximately 20 times more sensitive as a substrate for LIF than previously used radiolabeled substrates such as 3H-BAEE and 3H-TAME. Moreover, hydrolysis of 3H-BPVAME by partially purified LIF preparations was significantly inhibited by 10(-8) to 10 (-5) M of guanosine 3',5'-cyclic monophosphate (cGMP), further supporting the hypothesis that cGMP acts as a regulator of LIF activity. Inhibition of LIF-induced esterolysis was also provided by dibutyryl cGMP, but only at concentrations 10(-7) to 10(-5) M; 8-bromo cGMP and adenosine 3',5'-cyclic monophosphate were both ineffective. These results provide support for the use of 3H-BPVAME as a more selective substrate to detect esterase activity in LIF preparations than heretofore described and the possible development of a biochemical assay for the measurement of this lymphokine.
The effects of mild reduction on the structure and function of C3.
Authors: Isenman D E , Sundsmo J S , Cooper N R
Published: December 2022

Reduction and alkylation of C3 in nondenaturing buffers by 2 mM DTT (1 hr, 23 degrees C) (C3 R/A) results in a > 99% loss of C3 hemolytic activity. In contrast, sham alkylated protein, or protein allowed to reoxidize from the mildly reduced state, showed full activity. The loss in lytic activity correlated with the alkylation of approximately 6 liberated sulfhydryls, all of which arose from intrachain disulfides in the alpha polypeptide chain of C3. Data from CD experiments indicate that these bonds are essential for maintaining the native conformation of the protein. The loss of hemolytic activity of C3 R/A was neither due to its inability to be cleaved by C3 convertases nor to a dysfunction of the cleavage-induced labile membrane binding site as cells bearing C3 convertases firmly bound C3 R/A to an extent 80% that of native C3. The hemolytic defect in C3 R/A was found to result from its inability, once deposited on C42-bearing cells, to bind C5 and thereby participate in the formation of a C5 convertase. Unlike the effect on hemolytic activity, C3 R/A showed only a partial loss (3-fold) in its ability to bind to C3 receptors on monocytes. Taken together, these studies differentiate in molecular terms several of the functional sites in C3 and stress the significance of intra-alpha-chain disulfides in maintaining the conformation in activated C3 required for binding of C5.
A "nonidiotypic" inhibition of influenza-immune H-2-restricted CTL by an anti-T cell serum.
Authors: Effros R B , Hiserodt J C , Bonavida B
Published: December 2022

A xenogeneic serum directed at alloimmune CTL is shown to block H-2-restricted anti-viral CTL activity. Extensive absorption studies with targets and effector cells indicate that the blocking is at the level of the CTL and not the target cells. Inhibition is demonstrated for influenza-specific cytotoxicity operating in the context of 3 distinct H-2 haplotypes, suggesting that the serum is not directed at antigen-recognition structures. Kinetic studies on the addition of RAT to an ongoing 51Cr release assay further support the hypothesis that inhibition is not operating via the idiotypic region of the antigen receptor. We suggest that RAT is specific for CTL surface structures involved in nonrecognition events--possibly the lytic sites or a conserved region (idiotype-negative) of th antigen-binding receptor.
Monoclonal antibodies to HLA-A,B, and Ia-like antigens inhibit colony formation by human myeloid progenitor cells.
Authors: Fitchen J H , Ferrone S , Quaranta V , Molinaro G A , Cline M J
Published: December 2022

Hybridomas derived from the fusion of murine myeloma cells with splenocytes from mice immunized with human cultured lymphoid cells secreted monoclonal antibodies to human cell surface antigens. Serologic and immunochemical assays showed that 4 monoclonal antibodies (Ab Q2/47, Q2/61, Q2/70, Q2/80) recognize framework determinants of Ia-like antigens and 1 monoclonal antibody (Ab Q1/28) reacts with determinants expressed on the heavy chain of HLA-A,B antigens. Both anti-HLA-A,B and anti-Ia-like antigen monoclonal antibodies caused complement-dependent inhibition of granulocyte-macrophage colony formation by human bone marrow grown in soft agar. Mixing experiments excluded the possibility of an indirect effect on progenitor cells by lysis of auxiliary cells. These results indicate that human myeloid progenitor cells express HLA-A,B and Ia-like antigens.
Appearance of receptors for Macaca speciosa red blood cells on human fibroblasts transformed by simian virus 40.
Authors: Curry R A , Pellegrino M A , Saxton R E , Jensen F , Ferrone S
Published: December 2022

Receptors for monkey red blood cells (MRBC) that are expressed on subpopulations of human lymphoid cells are coded by genes linked to the major histocompatibility complex (MHC) region. Since malignant transformation of cells is associated with changes in structures coded by the MHC region, 10 cultured human melanoma and sarcoma cells and autologous SV40-transformed fibroblasts were tested for expression of MRBC receptors and compared with normal autologous fibroblasts. Only 2 of the tumor cell lines and normal fibroblasts from the same individual formed rosettes with MRBC. On the other hand, SV40 transformation induced in all the fibroblasts expression of receptors for MRBC. MRBC receptors on SV40-transformed fibroblasts show properties similar to those on B lymphoid cells.
Immunoglobulin synthesis by neoplastic B lymphocytes: free light chain synthesis as a marker of B cell differentiation.
Authors: Hannam-Harris A C , Gordon J , Smith J L
Published: December 2022

Immunoglobulin synthesis has been investigated in 55 cases of B lymphocytic disease. Free light chain was the predominant product of Ig synthesis and the sole secreted Ig product from neoplasms lacking surface Ig or expressing sparse surface IgM as detected by fluorescent-labeled antibody. In contrast, neoplastic cells expressing strong surface IgM or IgG synthesized and secreted balanced amounts of heavy and light chain. Intermediate patterns of synthesis were shown by neoplastic cells expressing IgM with IgD. These patterns of synthesis occurred irrespective of the disease entity and are discussed in relation to models of B cell maturation.
Monoclonal and allospecific H-2Kk antisera precipitate a high molecular weight protein.
Authors: Hunter F A , Paslay J W , Bhown A S , Mole J E , Bennett J C
Published: December 2022

A protein with an apparent m.w. of 68,000 has been observed in immunoprecipitations of NP-40 solubilized BW5147 thymoma cells by using monoclonal or polyclonal allospecific H-2Kk antisera. By contrast, H-2Dk alloantisera precipitated a 45,000 m.w. protein from the same membrane preparations. The 68,000 m.w. protein is expressed on the plasma membrane as determined by vectoral labeling. Extensive clearing experiments and the use of monoclonal antibody demonstrate that the protein does not bear group-specific C-type viral determinants.
Gangliotetraosyl-ceramide, the rat T lymphocyte-macrophage-associated antigen: chemical detection and cellular distribution.
Authors: Momoi T , Wiegandt H , Arndt R , Thiele H G
Published: December 2022

The presence of gangliotetraosyl-ceramide, i.e., Gal beta 1 leads to 3GalNAc beta 1 leads to 4Gal beta 1 leads to 4Glc beta 1 leads to Cer, the T lymphocyte-macrophage-associated antigen, on lymph node cells, thymic lymphocytes, and peritoneal exudate cells of the rat was established. For this, the glycosphingo-lipids of the cells were extracted and isolated by a microseparation and detection method. In contrast to immunoglobulin-negative rat lymph node cells, immunoglobulin-bearing lymph node cells showed no detectable gangliotetraosyl-ceramide.
Studies on the allergenic significance and structure of rat serum albumin.
Authors: Wahn U , Peters T , Siraganian R P
Published: December 2022

This study evaluated the capacity of rat serum albumin and of its proteolytic fragments to activate human basophils for IgE-mediated histamine release. The leukocytes from 8 out of 33 patients allergic to rats released histamine with rat serum albumin. Two proteolytic fragments of rat serum albumin, each constituting half of the molecule, were used to study the IgE-reactive antigenic sites. These fragments released histamine with the cells of some of the donors, thus demonstrating the presence of at least 2 antigenic determinants on each fragment for a total minimum of 4 sites on the intact rat serum albumin molecule. Most of the allergenic activity, however, was not recovered in the 2 fragments (total recovery mean = 6.4%, range between 0.1 and 31%). This loss could be due to cleavage of the rat albumin molecule in the middle of the third domain with loss of antigenic sites and/or due to minor conformational changes in the fragments as compared with the intact molecule. There was up to a 500-fold difference in the percent of activity recovered in the fragments when tested on cells from different patients. Therefore, there is no single immunodominant site on the molecule equally important for all patients. The cells of all 8 patients also reacted with mouse serum albumin but only 2 with bovine serum albumin. At least 1 determinant on mouse and rat serum albumin is cross-reactive with IgE.
IgG or IgM monoclonal antibodies reactive with different determinants on the molecular complex bearing Lyt 2 antigen block T cell-mediated cytolysis in the absence of complement.
Authors: Sarmiento M , Glasebrook A L , Fitch F W
Published: December 2022

Five rat monoclonal antibodies have been derived that express specificities for determinants present on the molecular complex bearing the Lyt 2 antigen. SDS-polyacrylamide gel electrophoresis of 125I-labeled polypeptides precipitated by each of these antibodies reveal 3 components (150,000, 75,000, and 33,000 daltons), and 2 components (44,000 and 33,000 daltons) when analyzed under nonreducing and reducing conditions, respectively. Two of these antibodies are IgG and are specific for the Lyt 2.2 determinant; the other 3 are IgM and react with determinants other than Lyt 2.2, which are nonpolymorphic. Each of the 5 antibodies can block the cytolytic activities of 5-day MLC cells or of cloned cytolytic T cells in the absence of C. Treatment of responding spleen cells with any of these antibodies and C inhibits the generation of cytolytic activity in MLC.
IgD allotypic determinants. I. Determinants expressed on murine IgD of the a allotype.
Authors: Woods V L , Kessler S W , Finkelman F D , Lieberman R , Scher I , Paul W E
Published: December 2022

Allotypes of membrane IgD of 27 strains of mice were determined with a series of anti-delta reagents, each capable of binding to the IgD of BALB/c spleen cells. One of these reagents is a newly defined allospecific rabbit anti-mouse-delta a. Typing with these reagents reveals the strain distribution of the previously described IgD.36 (Ig5.1) and IgD.43 (Ig5.4) specificities and demonstrates the existence of a new specificity, IgD.45 (Ig5.5). The results confirm the previous subdivision of the Igh-Ce haplotype into Igh-Ce, to which A mice are assigned, and Igh-Cn, to which NZB and NZW mice are assigned. In addition, it is shown that the Igh-Cd haplotype must also be subdivided. AKR mice, which express the a allele at the Igh-5(delta) locus, are designated as possessing the Igh-Cd haplotype while AL/N and C.AL20 mice, which have the e allele at the Igh-5(delta) locus, are assigned a new Igh-C haplotype designation, o.
Differences in cell-bound C8 sites on chicken erythrocytes measured by their reactivity with guinea pig and human C9.
Authors: Boyle M D , Gee A P , Okada M , Borsos T
Published: December 2022

Methods for preparing chicken erythrocytes (CE) with C7 bound to their surface were devised by using either the classical pathway (CEA1-7) or an activated 56hu reagent (CE567) derived from inulin-treated human serum. These intermediates were used to study the lysis of CE by functionally purified C8 and C9 isolated from guinea pig and human serum. The results indicated that GPC9 was less efficient in lysing CEA1-8 or CE5678 than HuC9. This finding was observed irrespective of the species of C8 used. Experiments designed to analyze this difference indicated that there were two functionally distinct forms of C5b-8 that were randomly distributed among the cells but differed in their ability to generate a C lesion depending on the species of C9 used to complete the reaction. The implications of these results on the mechanism of generation of C lesions are briefly discussed.
Further studies of the human platelet receptor for quinine- and quinine-dependent antibodies.
Authors: Kunicki T J , Russell N , Nurden A T , Aster R H , Caen J P
Published: December 2022

Previous studies have shown that the receptor for quinine- and quinidine-dependent antibodies is not expressed on the surface of platelets from patients with the Bernard-Soulier (B-S) syndrome. We now report data to suggest that these platelets lack the receptor for these antibodies. Since B-S platelets are also missing 2 related components of the GP I complex, GP lb and glycocalicin, our findings suggest that the receptor for quinine- and quinidine-dependent antibodies may be associated with this complex on normal platelets. An antibody previously shown to be directed against a surface antigen that migrated in the GP I position on SDS-polyacrylamide gel electrophoresis specifically blocked the reaction of the receptor with quinine- or quinidine-dependent antibodies. Purified glycocalicin, however, lacked detectable receptor activity. In contrast, a mixture of GP lb and a putative structural analog of this glycoprotein (Mr 210,000) eluted from wheat germ affinity columns after chromatography of Triton X soluble preparations of platelets or membranes was shown to contain at least 80% of the total receptor activity. Our results strongly suggest that the receptor is associated with GP lb and/or its high m.w. structural analog(s).
Differential effect of aging on the heterogeneity of the immune response to a T-dependent antigen in systemic and mucosal-associated lymphoid tissues.
Authors: Szewczuk M R , Campbell R J
Published: December 2022

The heterogeneity of the immune response in various lymphatic sites to trinitrophenylated bovine gamma-globulin was measured in old and young mice at a cellular level using inhibition of plaque-forming cell assay. The primary splenic PFC response of old C57BL/6J male mice was found to be markedly restricted in heterogeneity, with a preferential loss of IgM, IgG, and IgA high avidity PFC. The draining peripheral lymph node IgG PFC response of old mice exhibited similar restriction in heterogeneity, with a preferential loss of high avidity PFC. In contrast, mesenteric and mediastinal lymph node PFC response of old mice remained highly heterogeneous with respect to antibody affinity in the 3 isotypes studied. These results support the hypothesis that the mucosal-associated lymphoid system differs from the systemic system with regard to its immune competence with age. Furthermore, the results imply that the T cell helper activity is unimpaired in the mucosal-associated lymphoid apparatus in aged animals.
Orientation of PMN in a pH gradient: acid-induced release of a chemotactic factor.
Authors: Zigmond S H , Hargrove R L
Published: December 2022

Human and rabbit PMN, placed in a gradient of pH 5 to pH 8 over 1 mm, orient their locomotion toward pH 5. This orientation is due to the release or production of a chemotactic factor by the cells in the acid pH. This activity can be recovered in the supernatant and is maximal when the cell incubation is at pH 5.5. Its appearance is rapid, occurring within 1 min, and is temperature dependent. Cells survive treatment at pH 5.5 and are capable of subsequently responding to chemotactic peptides.
Fate of a common acute lymphoblastic leukemia antigen during modulation by monoclonal antibody.
Authors: Pesando J M , Ritz J , Lazarus H , Tomaselli K J , Schlossman S F
Published: December 2022

Modulation of a human common acute lymphoblastic leukemia antigen (CALLA) by specific monoclonal antibody (J5) has been studied with the immune precipitation method to identify radiolabeled antigen. Surfaces of leukemic cells have been labeled using 125I both before and after modulation by J5 antibody for different time intervals. Leukemic cells have also been metabolically labeled with 35S-methionine before modulation. These studies indicate that the 100,000-dalton glycoprotein expressing CALLA (gp 100-CALLA) cannot be detected in cells that were modulated with J5 antibody before surface labeling but that it is easily detectable in cells that were surface labeled before modulation for 10 hr. At later time points, gp 100-CALLA is selectively lost from cells that were surface labeled before modulation. Gp 100-CALLA is not detected in the supernatants from cultures of these modulated cells. We conclude that gp 100-CALLA is rapidly internalized during modulation and that CALLA is degraded. Gp 100-CALLA is not shed into the culture media, nor does it remain on the cell surface in an altered form. Incubation of leukemic cells with antisera to beta2-microglobulin or IgM does not affect the expression of gp 100-CALLA.
Characterization of a novel human pre-B leukemia cell line.
Authors: Smith R G , Dev V G , Shannon W A
Published: December 2022

The derivation of a new human pre-B cell line from the leukocytes of a girl in the leukemic phase of lymphoblastic lymphoma is reported. Cells of this line, termed SMS-SB, synthesized mu but not light chains. These mu-chains were found only in the cytoplasm and were not secreted. SMS-SB cells bore HLA-DR determinants. They did not express the common ALL antigen or terminal deoxynucleotidyl transferase and, therefore, differed from previously described human pre-B leukemia cell lines. SMS-SB cells resembled certain Abelson leukemia virus-transformed mouse bone marrow cells. Except for the lack of formed mouse bone marrow cells. Except for the lack of mu secretion, the phenotype of SMS-SB cells was the same as the major subpopulation of marrow pre-B cells.
Fc fragment activation of T lymphocytes. I. Fc fragments trigger Lyt 1+23- T lymphocytes to release a helper T cell-replacing activity.
Authors: Thoman M L , Morgan E L , Weigle W O
Published: December 2022

Fc fragments of human IgG stimulate both proliferation and polyclonal antibody formation by B lymphocytes. T lymphocytes, although not proliferating in response to Fc fragments, are triggered to produce a T cell-replacing factor, which substitutes for T cells in the Fc fragment-induced polyclonal response. The factor is produced within 24 hr after Fc stimulation. Neither Fab fragments nor whole IgG have the capacity to stimulate the release of this activity. This material is a product of Lyt 1+23-, Ia-T lymphocytes and requires Ia+ adherent accessory cells for production. The role of the accessory cell is to process the Fc fragments to biologically active 14,000-daltons subfragments, which directly trigger T cells to factor production. This material has been called (Fc)TRF to distinguish it from other T cell factors.
Analysis and antigenic activity of a carbohydrate fraction derived from adult Schistosoma mansoni.
Authors: Nash T E , Lunde M N , Cheever A W
Published: December 2022

The antigenic constituents of a crude predominately carbohydrate trichloroacetic acid soluble fraction of adult Schistosoma mansoni (TCA-S-C) were determined by comparing the antibody levels in human infections with crude TCA-S-C and fractions obtained after DEAE chromatography. Antibodies were measured using ELISA tests employing poly(L-lysine)-coated and uncoated polystyrene plates as well as radioimmunoassays to the 2 active fractions. Two active materials were identified. Acute and early infected patients had high levels of antibody to the previously characterized purified gut-associated proteoglycan (GASP), whereas the heavily infected chronically exposed patients had elevated levels to another fraction, PSAP. IgG levels to PSAP correlated significantly with the egg excretion rate. Both the acute and early infected as well as the chronically exposed patients had raised levels of antibody to crude TCA-S-C but no correlation with egg excretion or acuteness of infection. Therefore, the measurement of antibodies to specific schistosome components unmasked clinically relevant correlations that were not apparent when crude fractions were used. The pattern of response was similar whether ELISA or radioimmunoassays were used, although the radioimmunoassays were found to be more sensitive and specific. Antibody levels to GASP significantly correlated with the immunofluorescent antibody titer to schistosome gut epithelial cells. PSAP was characterized as a polydisperse PAS-positive staining material composed primarily of substances(s) with an apparent m.w. ranging from 130,000 to 90,000. The composition of this material as revealed by earlier studies suggested that PSAP was a glycoprotein(s).
Regulation of the immune response: III. The role of macrophages in the potentiation of the immune response by Fc fragments.
Authors: Morgan E L , Weigle W O
Published: December 2022

The Fc fragment-induced enhancement of the anti-SRBC response was found to be dependent upon the presence of functional macrophages. Fc fragments derived from human immunoglobulin were unable to enhance either the in vivo or in vitro anti-SRBC response in the C3H/HeJ mouse, whereas similar responses in histocompatible C3Heb/FeJ mice were enhanced. Macrophages were found to be required for the Fc fragment-induced adjuvant effect as evidenced by the inability of C3H/HeJ macrophages to substitute for C3Heb/FeJ macrophages in Sephadex G-10-filtered C3Heb/FeJ spleen cell cultures. Moreover, the C3H/HeJ anti-SRBC response could be enhanced when Fc subfragments were employed instead of Fc fragments. The C3H/HeJ-derived T cells were normal, since they could be added to B cells and macrophages derived from the C3Heb/FeJ mouse with an enhanced response observed in the presence of Fc fragments. The fact that C3H/HeJ macrophages function normally as accessory cells in generation of the anti-SRBC response but fail to function in the Fc-induced adjuvant effect allows the dissection of 3 macrophage functions in the modulation of the antibody response.
Clonal variation in expression of a human melanoma antigen defined by a monoclonal antibody.
Authors: Yeh M Y , Hellström I , Hellström K E
Published: December 2022

Previous work established 3 monoclonal mouse antibodies, 3.1, 3.2, and 3.3, which define an antigenic determinant that is expressed by a human melanoma, M1804, and, in smaller amounts, by another melanoma, M1801. In the present study we first investigated the expression of this determinant, which we refer to as 3.1, by cells from different melanomas, using membrane immunofluorescence techniques. We found that some tumors, such as M1801, appeared to be mixtures of cells that varied in their expression of 3.1. Clones were established from cultures of M1801 and M1804, and their expression of 3.1 was studied by a variety of serologic techniques, including membrane immunofluorescence, complement-dependent cytotoxicity, and absorption. Some of the clones expressed determinant 3.1, and others did not. By recloning a late passage of a 3.1-positive clone from M1801, a negative subclone was established, proving that 3.1-positive cells can yield 3.1-negative progeny. A clone that had lost 3.1 was tested and found to still express p97, an antigen defined by a different monoclonal antibody. We conclude that since some melanomas are heterogeneous with respect to the expression of antigenic determinant 3.1, and since 3.1 negative progeny can be derived from 3.1-positive cells, there is the potential for selection of 3.1-negative cells. The implications of this, for antigenic determinant 3.1 and possibly for other tumor antigens, must be taken into account when considering using monoclonal antibodies for tumor therapy.
Cell surface appearance of unexpected host MHC determinants on thymocytes from radiation bone marrow chimeras.
Authors: Sharrow S O , Mathieson B J , Singer A
Published: December 2022

The phenotypic appearance of cell surface antigens on murine thymocytes from long-term radiation bone marrow chimeras was analyzed using indirect immunofluorescence and flow microfluorometry. Cells maturing in the thymi of these mice were typed for MHC (Kk, I-Ak, H-2b, Kb, and Ib) and non-MHC (Lyt 1, Ly 9, and TL) determinants. All cells were of donor origin as determined by non-MHC (Ly) phenotype in P1 leads to P2, P1 X P2 leads to P1, and P1 leads to P2 radiation chimeras. In contrast, the MHC phenotypes of these thymocytes were markedly affected by the host environment. Specifically, H-2 and I-A determinants of both parental phenotypes were detected on thymocytes from P1 leads to P1 X P2 chimeras; I-A determinants of host phenotype were present, whereas I-A determinants of donor phenotype were reduced on thymocytes from P1 X P2 leads to P1 chimeras; and thymocytes from P1 leads to P2 chimeras possessed H-2 and I-A determinants of host phenotype but showed reduction of donor I-A phenotype determinants. The appearance of host cell surface H-2 and I-A determinants on thymocytes from chimeras closely parallels the functional recognition of MHC determinants by T cells from chimeric mice and thus may be significantly related to the development of the self-recognition repertoire by maturing T cells.