bluuebunny/crossref_metadata_2025 huggingface dataset:
Functional properties of lymphocyte subpopulations in hepatitis B virus infection. II. Cytotoxic effector cell killing of targets that naturally express hepatitis B surface antigen and liver-specific lipoprotein.
Authors: Chisari F V , Bieber M S , Josepho C A , Xavier C , Anderson D S
Published: December 2022

Cytotoxic effector cell responsiveness to host and/or virus-determined hepatocyte surface membrane antigens has been postulated as an important pathogenetic determinant of hepatocellular injury in hepatitis B virus infection. Assuming that such effector cell populations would be detectable in peripheral blood, the present study was designed to examine 2 questions: first, whether target cells that normally express liver-specific protein (LSP) and hepatitis B surface antigen (HBsAg) are selectively destroyed by peripheral blood effector cells from patients with viral hepatitis; second, whether cytotoxic effector cell function emerges coincident with the development of defective suppressor cell activity in the same patients. No evidence of increased HBsAg or LSP specific cytotoxic effector cell activity was found in the peripheral blood natural killer (NK) or T killer cell populations of patients with acute or chronic viral hepatitis.
Binding of bacterial endotoxin (LPS) to encephalitogenic myelin basic protein and modulation of characteristic biologic activities of LPS.
Authors: Raziuddin S , Morrison D C
Published: December 2022

Myelin basic protein, isolated from central nervous system tissue and an inducer of experimental allergic encephalomyelitis in animals, has been demonstrated to form a stable molecular complex with the lipid A region of gram-negative bacterial lipopolysaccharides (endotoxins). This binding of endotoxin with myelin basic protein results in generation of lower m.w. aggregates with decreased isopycnic density. A number of lipid A-induced characteristic properties of endotoxin, such as B lymphocyte proliferative response in C3H/St mice, complement activation of normal human serum, Limulus lysate gelation, and lethal effects in mice, are modified as a result of binding of myelin basic protein with lipopolysaccharides.
Suppression of anti-hapten IgE and IgG antibody responses by isologous anti-idiotypic antibodies against purified anti-carrier (ovalbumin) antibodies in BALB/c mice.
Authors: Blaser K , Nakagawa T , de Weck A L
Published: December 2022

The studies described here show that BALB/c mice actively producing isologous anti-idiotypic antibodies (ald) after immunization with purified, syngeneic anti-OvA antibodies exhibit depressed anti-hapten and anti-carrier IgE and IgG antibody responses, provided that hapten-OVA conjugates are used to mount the immune responses. We suggest that carrier-specific helper T cells (CTh) for IgE and IgG antibody production carry idiotypes similar or identical to those among purified anti-OVA IgG antibodies. The mode of regulation of immune responses by anti-carrier-ald may be different from that exerted by anti-hapten-alD. Although anti-carrier-ald may block or deplete CTh and thereby prevent help for immunoglobulin production by B cells, anti-hapten-ald may induce T suppressor cells (Ts).
Antibody precipitation of lipid vesicles containing myelin proteins: dependence on lipid composition.
Authors: Boggs J M , Clement I R , Moscarello M A , Eylar E H , Hashim G
Published: December 2022

Antibody binding to human CNS myelin basic protein and to rabbit sciatic nerve myelin P-2 in their lipid-bound and water-soluble conformations has been investigated. 125I-labeled basic protein or P-2 was bound to the surface of liposomes (vesicles) of different acidic lipids, phosphatidylethanolamine (PE), phosphatidylserine (PS), phosphatidic acid (PA), phosphatidylglycerol (PG), and cerebroside sulfate (CBS). The antibody was prepared against aqueous solutions of basic protein and P-2. Antibody binding to the proteins in liposomes was measured by precipitation of the liposomes by using a double antibody radioimmunoassay. The amount of 125I-basic protein precipitated was lest when the protein was bound to PA and increased in the order PA less than PS less than PG less than CBS less than PE approximately equal to basic protein in solution, suggesting that the antigenic determinants were lest exposed or most altered for PA and most exposed for PE. This agreed fairly well with previously published biophysical studies that suggested that hydrophobic segments of the protein penetrated into the lipid bilayer and that this penetration decreased in the order PA approximately equal to PG greater than PS greater than CBS greater than or equal to PE. The amount of 125I-P-2 precipitated was least for PA and CBS and increased in the order PA approximately equal to CBS less than PS less than PG less than PE approximately equal to P-2 in solution. However, the differences were less than for basic protein and the effect of CBS was different for the 2 proteins. Less is known about the conformation of P-2 in these lipids but it is known that lipids increase its disease-inducing activity. These results indicate that interaction with lipid may sequester or alter the conformation of antigenic determinants such that antibody binding decreases.
Agglutination of complement-coated erythrocytes by serum amyloid P-component.
Authors: Hutchcraft C L , Gewurz H , Hansen B , Dyck R F , Pepys M B
Published: December 2022

Serum amyloid P-component (SAP) is a normal plasma glycoprotein apparently identical with the P-component associated with amyloid deposits. SAP shows extensive amino acid sequence homology with the C-reactive protein, and both have a similar molecular configuration. SAP undergoes calcium-dependent binding to zymosan, agarose, and amyloid fibrils, but its functional properties are not yet known. We report here that SAP agglutinates complement-(C) coated antibody-sensitized erythrocytes by a calcium-dependent reaction. SAP was found to interact predominantly with a modified form of bound C3b. This modification was achieved by prolonged treatment of EAC43 with heated normal human serum or with isolated C3bINA and beta 1H, and reactivity was reduced upon treatment of the cells with trypsin. SAP thus seems to react with fixed C3 in a manner similar to the reaction of C3 with bovine conglutinin, a molecule that also undergoes calcium-dependent binding to zymosan and agarose. These studies identify a new reactivity for SAP and demonstrate an interaction between an amyloid protein and the C system. The close similarity between the calcium-dependent binding specificities of SAP and of bovine conglutinin may assist in characterization of these molecules and in investigation of their function.
The production of anti-IgG2a autoantibody in the 129/Sv mouse: onset in the lymph nodes draining the intestinal tract and prevention by neonatal thymectomy.
Authors: Van Snick J L
Published: December 2022

The histological distribution of anti-IgG-secreting cells was studied by measuring the amount of anti-IgG autoantibodies produced in vitro by cells isolated from various lymphoid organs. Before 20 wk, the production of anti-IgG autoantibody in 129/Sv mice from our Institute predominated in the mesenteric and caudal lymph nodes, which both drain the intestinal tract. After 20 wk, the production extended to the spleen and bone marrow. At that time no production was detected in the mediastinal, lumbar, and inguinal lymph nodes. A similar distribution of anti-IgG-secreting cells was observed in 25-wk-old 129/J mice from the Jackson Laboratory. These data indicate that anti-IgG autoimmunity in the 129 primarily results from a local immune response, probably induced by a specific stimulus of intestinal origin. Neonatal thymectomy prevented the production of anti-IgG, demonstrating that the mesenteric anti-IgG production did not result from the direct stimulation of B lymphocytes by endotoxin-like polyclonal activators. It was found that the anti-IgG produced in vitro had the same narrow specificity against IgG2a as its serum counterpart, indicating that this specificity resulted from the selective stimulation of anti-IgG2a-secreting cells rather than from in vivo consumption of anti-IgG antibodies recognizing other IgG subclasses.
Trypanosome variable surface antigens: studies using two-dimensional gel electrophoresis and monoclonal antibodies.
Authors: Pearson T W , Kar S K , McGuire T C , Lundin L B
Published: December 2022

Variable surface antigens of cloned populations of African trypanosomes were studied using 2-dimensional gel electrophoresis and monoclonal antibodies. Two-dimensional gel maps showed that the only major differences in protein profiles of the non-nuclear materials from different clones of solubilized trypanosomes were attributable to the variable surface antigens and that purification doesn't alter these molecules, at least with respect to charge and apparent m.w. Purified variable surface antigens were used as immunogens and a number of monoclonal antibodies were derived using cell-fusion technology. In radioimmunoassay, each of the monoclonal reagents was shown to be specific for the immunizing antigen, and in immunofluorescence tests using acetone-fixed trypanosomes, each bound specifically to the clone from which the antigens were purified. Only 20% of the monoclonal reagents bound to living trypanosomes, however, providing evidence for both exposed and nonexposed antigenic sites on the variable surface antigens. Those reagents derived to a single antigen bound to different nonoverlapping antigenic sites, which were on the protein portion of the molecules and not the carbohydrate moieties. The data show that monoclonal antibodies are ideal probes for studying localization of antigenic sites on the antigen molecules and for studying antigen synthesis, glycosylation, and architecture in relation to the cell surface. In addition, the finding of exposed and nonexposed antigenic sites on trypanosome variable antigens allows a possible explanation for the role of the antigens in pathogenesis of African trypanosomiasis.
Stimulation of human synovial fibroblast plasminogen activator production by mononuclear cell supernatants.
Authors: Hamilton J A , Slywka J
Published: December 2022

Conditioned medium from concanavalin A-stimulated human peripheral blood mononuclear cells (c-MCCM) stimulates the plasminogen activator (PA) production of nonrheumatoid human synovial fibroblasts obtained from explant cultures. The effect of this synovial fibroblast-stimulating activity is observed within 2 to 4 hr and requires RNA and protein synthesis. Reversible morphological changes in the synovial cells can be observed as a result of c-MCCM action. These enzymatic and morphologic changes are similar to some of the effects of transforming viruses and tumor promoters on target cells. The possible significance of these data for an understanding of the cellular interactions involved in the formation and function of the rheumatoid "pannus" is discussed.
Interaction of C-reactive protein with liposomes. III. Membrane requirements for binding.
Authors: Mold C , Rodgers C P , Richards R L , Alving C R , Gewurz H
Published: December 2022

We have previously presented a model for CRP-membrane interactions using liposomes composed of dimyristoyl phosphatidylcholine (DMPC), cholesterol (CHOL), stearylamine (SA), and galactosyl ceramide. We have shown that the interaction of cRP with these liposomes in the presence of human serum results in consumption of hemolytic C and membrane lysis. In the present paper we have directly examined the binding reaction between CRP and liposomes using radiolabeled CRP. We have found that this binding is more characteristic of CRP interactions with polycations than CRP interactions with phosphocholine- (PC) containing molecules. CRP binding to liposomes was dependent on the presence of SA in the membrane and could occur with dimyristoyl phosphatidylethanolamine in place of DMPC. The binding was not inhibited by ethylenediaminetetraacetate (EDTA) but could be inhibited by CaCl2, whereas CRP binding to PC-Sepharose was inhibited by EDTA and required CaCl2. We have further examined the effects of changes in membrane composition on CRP binding to liposomes. In liposomes with a limiting density of SA, we found increased CRP binding with changes in composition that would increase membrane fluidity. In most cases, the amount of CRP binding correlated with the amount of C activation observed previously. However, increasing the amount of CHOL in the membrane was found to increase C activation while decreasing CRP binding. These findings indicate that CRP binding to membranes and subsequent C activation can occur through cationic molecules as well as phospholipids.
Relative lack of Epstein Barr virus (EBV) receptors on B cells from persistently EBV seronegative adults.
Authors: Gervais F , Wills A , Leyritz M , Lebrun A , Joncas J H
Published: December 2022

Viral receptors are essential for the entry of the virus into the cell. EBV receptors can be detected on fresh lymphocytes by a technique that uses EBV-coupled tanned red blood cells that form rosettes with lymphocyte-bearing receptors. This technique was found to detect viral receptors only and not surface immunoglobulins. T cell depletion of the lymphocyte population showed that these receptors were present on B lymphocytes. Study of the presence of these EBV receptors on the surface of fresh lymphocytes from 66 subjects (age 2 to 66), selected out of a group of over 2000 individuals, showed that the majority of these donors had receptors for the virus. However, a few of these adults persistently failed to develop anti-EBV antibodies, even if they were in close contact with the infectious agent. The lymphocytes of 11 such individuals were found to be lacking EBV receptors. Transformation assay of these lymphocytes did not give rise to lymphoblastoid cell lines whereas lymphocytes from 4 individuals, who were EBV seropositive or seronegative but receptor positive, yielded permanent lymphoblastoid cell lines. This would suggest that a few EBV seronegative adults (less than 0.5%) display natural resistance to EBV transformation of their lymphoid cells as a result of absolute or relative lack of EBV receptors on these cells.
Mycoplasma-mediated activation of normal mouse lymphocytes: induction of cytotoxic lymphocytes and lymphocyte transformation by Mycoplasma arthritidis are under Ir gene control.
Authors: Cole B C , Daynes R A , Ward J R
Published: December 2022

The ability of M. arthritidis, a potent arthritogenic agent, to induce normal unsensitized lymphocytes to undergo blast transformation and to become nonspecifically cytotoxic for syngeneic and allogeneic target fibroblasts was found to be dependent upon the donor strain of mouse. Lymphocytes from CBA (H2k) and BALB/c (H2d) mice were identified as high responders and lymphocytes from C56BL/10 mice (H2b) as low or nonresponders. In the cytotoxicity assay the extent of cell lysis was dependent upon the origin of both the effector cell and the target cell. By comparing the ability of M. arthritidis to activate lymphocytes from various congenic and H-2 recombinant mouse strains we demonstrated that control of the degree of responsiveness was determined by the H2 gene complex. Further studies revealed that the region controlling induction of cytotoxic lymphocytes is located to the right of the I-J subregion and to the left of the D region, most probably the I-E and/or I-C subregions of the major histocompatibility complex. These observations may be of importance in regard to the susceptibility of different mouse strains to the pathogenic effects of M. arthritidis.
Correlation between clinical activity of systemic lupus erythematosus and the amounts of DNA in DNA/anti-DNA antibody immune complexes.
Authors: Morimoto C , Sano H , Abe T , Homma M , Steinberg A D
Published: December 2022

The relationship between clinical activity of systemic lupus erythematosus (SLE) and molecular sizes of DNA fragments isolated from DNA/anti-DNA antibody immune complexes were examined. Among sera from twenty-eight patients with SLE examined, three different molecular sizes of DNA were identified, namely, DNA of m.v. 25,000 with 30 to 50 base pairs (bp); m.w. 100,000 with 150 to 200 bp; and m.w. 200,000 with 300 bp. On the basis of the molecular sizes of DNA fragments, we can divide patients with SLE into four groups. The first group contained DNA predominantly of m.w. 25,000. The second group contained DNA predominantly of m.w. 100,000. The third group contained DNA of m.w. 200,000. The fourth group contained both m.w. 25,000 and 100,000. A method to quantitate the amounts of DNA in DNA/anti-DNA immune complexes was developed. The amount of DNA fragments was estimated by measuring the amount of 32P-phosphate incorporated into 5 ends of DNA. Patients with severe disease tended to have greater amounts of DNA (up to 400 ng/ml serum). Two serial studies also support this result. Thus, the quantitative analysis shows that the amount of DNA in the immune complexes is highly correlated with disease activity (r = 0.864; p less than 0.001). These results suggest that DNA/anti-DNA immune complexes may play some role in the pathogenesis of lupus nephritis.
Oligoclonal IgG bands with and without measles antibody activity in sera of patients with subacute sclerosing panencephalitis (SSPE).
Authors: Mehta P D , Patrick B A , Thormar H , Wisniewski H M
Published: December 2022

Oligoclonal IgG bands from SSPE sera were isolated by combination of Protein A-Sepharose 4B column and preparative isoelectric focusing gel procedures. Each eluted fraction, when examined in analytic IEF, showed two or three individual bands with isoelectric points close to one another, compared to approximately fifteen IgG bands seen in whole serum. When the bands were tested for measles antibody activity in immunofixation with measles virus followed by peroxidase staining, the bands eluted in pH region 8.5 to 9.3 were found to be measles specific, whereas those in pH 7.0 to 8.4 lacked significant measles activity. When eluted fractions containing groups of bands were absorbed with measles virus, the bands in pH region 8.5 to 9.3 were removed, whereas those in pH 7.0 to 8.4 region remained unchanged; this indicated that a number of oligoclonal IgG bands without measles virus activities are present in SSPE. The bands lacking measles-specific activity may be synthesized against other infectious agents or they may represent nonspecific activation of B cell clones.
Anti-idiotypes to monoclonal anti-H-2 antibodies. III. Syngeneic anti-idiotypes on antibodies induced by in vivo administration of xenogeneic anti-idiotypes.
Authors: Bluestone J A , Auchincloss H , Cazenave P A , Ozato K , Sachs D H
Published: December 2022

Idiotype-positive molecules induced by xenogeneic anti-idiotypes (anti-Id) to monoclonal anti-H-2 antibodies were studied using syngeneic anti-Id reagents. Syngeneic reagents detected idiotypes on both H-2 antigen-binding and nonbinding Id' induced by xenogeneic anti-Id. In addition, idiotopes were readily detected by alloimmunization after xenogeneic anti-Id reagents interact with some of the same antibodies as syngeneic anti-Id and support the hypothesis that treatment with xenogeneic anti-Id may affect a physiologically important idiotypic network.
Virulent P. berghei malaria: prolonged survival and decreased cerebral pathology in cell-dependent nude mice.
Authors: Finley R W , Mackey L J , Lambert P H
Published: December 2022

The course of lethal Plasmodium berghei infection was examined in nu/+ and T cell-deficient nu/nu BALB/c mice. A rapidly fatal neurologic syndrome, including ataxia, hemiparesis, and seizures, was seen in the nu/+ mice early in the infection, whereas this syndrome was absent in the nu/nu mice. The nu/nu mice also developed anemia more slowly, had lower levels of immune complexes and total IgG, and had smaller decreases in serum C3 compared with the nu/+ mice. Histopathologic examination of the brains revealed cerebral malaria lesions, including vascular plugging and micro-hemorrhages, in the nu/+ mice but not in the nu/nu mice. Cerebral lesions similar in frequency and severity to those in nu/+ mice developed in nu/nu mice given spleen cells from normal nu/+ mice. The results suggest that an intact immune system is necessary for the expression of cerebral malaria.
Purification and characterization of a high molecular weight eosinophil chemotactic factor from Schistosoma japonicum eggs.
Authors: Owhashi M , Ishii A
Published: December 2022

A high m.w. eosinophil chemotactic factor (ECF-SjE) was isolated and purified from a soluble egg antigen preparation (SEA) of Schistosoma japonicum by gel filtration on Sephacryl S-200, anion-exchange chromatography on DE52, and isoelectric focusing. ECF-SjE had a m.w. of more than 900,000 and an isoelectric point of 4.1. It contained 40% (w/w) sugar residues and bound to concanavalin A (Con A). The chemotactic activity of ECF-SjE was heat stable (100 degrees C, 60 min) and resistant to pronase digestion, but was destroyed by periodate oxidation. IgG antibody to ECF-SjE was detected in the serum of a rabbit infected with S. japonicum, demonstrating the antigenic nature of ECF-SjE. The antigenicity of ECF-SjE was also sensitive to periodate oxidation. Thus, ECF-SjE is a glycoprotein or proteoglycan from the eggs of S. japonicum, and the sugar chain is important for the expression of chemotactic and antigenic activities. However ECF-SjE differs from the major allergenic components of S. japonicum (JEAL) in m.w. and isoelectric point. A low m.w. eosinophil chemotactic factor was also detected in SEA. Together they are proposed to have a role in the direct accumulation of eosinophils in the egg-induced granulomas in S. japonicum infection.
Monoclonal antibodies to human alpha-interferon and their use for affinity chromatography.
Authors: Novick D , Eshhar Z , Rubinstein M
Published: December 2022

A new screening procedure has been developed and used for the identification of three hybridomas secreting monoclonal antibodies to human leukocyte interferon (IFN-alpha). The screening procedure is based on immune precipitation with a second antibody, acid dissociation of the precipitate, and bioassay of the recovered interferon. This procedure selects for monoclonal antibodies that are suitable for affinity chromatography, as demonstrated by construction of immunoadsorbents from these monoclonal antibodies and purification to homogeneity in IFN-alpha.
Results from the Use of the Blood Clot Method of Immunization
Authors: Anderson John F. , Leonard George F.
Published: December 2022

Zinsser and Grinnell in 19252 reported a novel method for the immunization of horses against pneumococcus, and suggested its application in the case of other bacteria. When this procedure came to our attention, we were immediately interested in the possibility of its use, particularly in the immunization of horses against certain types of streptococci, pneumococci and other bacteria. We have used the method for the immunization of horses against different strains of streptococci, pneumococci, the green producing diplococcus isolated by Tunnicliff from cases of measles, and a combination of B. coli and streptococci.

The essential feature in the blood clot method is the injection of whole oxalated blood to which the bacteria have been added, either immediately before injection or sometime previous, together with sufficient calcium chloride to induce the formation of a clot within a few minutes. It was the idea of the originators that the toxic factor from the pneumococcus developed when the pneumococci grew in the blood under conditions analogous to those prevailing in pneumonia, although they were undecided whether this toxic factor was antigenic.
Interferon-gamma induces class I HLA and beta 2-microglobulin expression by human amnion cells.
Authors: Hunt J S , Wood G W
Published: December 2022

In contrast to other human nucleated cells, amnionic epithelial cells do not normally express class I HLA antigens. It is not known whether the failure of display of those antigens occurs because genes for HLA expression are deleted or are defective, or if gene expression is repressed. The results of the present study demonstrate that amnionic epithelial cells exposed to IFN-gamma express levels of class I HLA antigens and beta 2-microglobulin that are readily detectable in immunoperoxidase and immunofluorescence stains. The findings suggest that failure of transcription may be responsible for failure of normal expression of class I HLA antigens and beta 2-microglobulin by cells of the amnion membrane. Class II antigens, which are normally expressed by very few cell types, were not expressed by amnion cells under any test conditions.
Reorientation and fusion of cytotoxic T lymphocyte granules after interaction with target cells as determined by high resolution cinemicrography.
Authors: Yannelli J R , Sullivan J A , Mandell G L , Engelhard V H
Published: December 2022

The interaction of murine cytotoxic T lymphocyte (CTL) clones with human lymphoblastoid target cells was studied in thin preparations by using high resolution cinemicrography. CTL not bound to target cells were morphologically polar, possessing a broad leading edge containing the nucleus, and a tapered tail containing a large number of granules. The CTL were observed to move by the extension of pseudopods from the leading edge. Initial contact with a target cell was made via the leading edge of the CTL. If the human target cell expressed the appropriate HLA antigen, distinct morphologic changes occurred in the CTL as early as 2 min after initial contact. The CTL rounded up, and the nucleus moved from a position adjacent to the zone of contact to be replaced by the cytoplasmic granules. Redistribution of the granules was completed as early as 10 min after initial contact was made. These morphologic changes did not occur when the CTL made contact with other CTL, or with target cells that did not express the appropriate HLA antigens. In studies that make use of Nomarski optics, an apparent fusion of CTL cytoplasmic granules with the membrane in the vicinity of the target cell contact area was observed 4 min after binding, and before granule reorientation was complete. These data provide direct evidence for the occurrence of both reorientation of the cytoplasmic contents and granule fusion in CTL with a time course similar to that of administration of the lethal hit.
Clonal analysis of the anti-Qa-1 cytotoxic T lymphocyte repertoire: definition of the Qa-1d and Qa-1c alloantigens and cross-reactivity with H-2.
Authors: Aldrich C J , Jenkins R N , Rich R R
Published: December 2022

To characterize the four common Qa-1 allelic products, we examined in detail the CTL-defined determinants encoded by Qa-1. In previous studies with anti-Qa-1 CTL and alloantisera, investigators have described antigenic determinants present on Qa-1a and Qa-1b antigens, but they have defined Qa-1c and Qa-1d exclusively by their cross-reactivity with Qa-1a and/or Qa-1b determinants. To delineate further the CTL-defined determinants encoded by Qa-1d, we generated CTL clones with Qa-1d specificity and demonstrated that the Qa-1d molecule expressed determinants that were not detected on Qa-1a, Qa-1b, or Qa-1c target cells. Other CTL clones derived from anti-Qa-1d MLC recognized new antigenic determinants on Qa-1c that cross-reacted with Qa-1d. Each of the four common Qa-1 phenotypes was shown to exhibit unique antigenic determinants. In addition, Qa-1d anti-Qa-1a and Qa-1d anti-Qa-1b CTL confirmed extensive cross-reactivity among these Qa-1 alloantigens. Analysis of CTL from these four immunizations also resulted in the isolation of Qa-1a-specific and Qa-1d-specific CTL clones that cross-reacted with H-2Df and H-2Ks, respectively.
Ia-positive nonlymphoid cells and T cell development in murine fetal thymus organ cultures: monoclonal anti-Ia antibodies inhibit the development of T cells.
Authors: DeLuca D
Published: December 2022

A fetal thymus organ culture system has been developed to study the differentiation of murine thymus-derived immunocompetent cells (T cells) such that cell yields can be easily monitored. This system has been used to study the effects of monoclonal anti-I-A antibodies on the growth of T cells. The addition of anti-I-A antibodies, but not anti-H2K monoclonal antibodies, to fetal thymus organ cultures resulted in a decreased yield of lymphoid cells. Anti-I-A-treated cultures did not produce cells that gave an immune response in MLC assays. Anti-I-A antibodies stained a small subpopulation of nonlymphoid cells in untreated cultures by indirect immunofluorescence that were no longer detectable in cultures that had been pretreated with anti-I-A antibody. Culture of fetal thymus lobes at low temperature (20 degrees C) for 1 wk resulted in a decrease in lymphocyte production, as well as a concomitant increase in the frequency of Ia-positive nonlymphoid cells. Co-culture of fetal liver or anti-thy-1 plus complement-treated adult bone marrow with such Ia-positive cell-enriched fetal thymus lobes at 37 degrees C resulted in the production of T cells. Anti-Thy-1.1 or -1.2 staining by indirect immunofluorescence of cells obtained from co-cultures that differed at the Thy-1 locus showed that the T cells produced were derived from the bone marrow or fetal liver. T cell production occurred in both syngeneic and allogeneic cocultures. However, if co-cultures were made by using 14-day gestation fetal thymus instead of fetal liver or bone marrow as donors of T cell precursors, T cell growth was observed only in syngeneic combinations. These results suggest that Ia-positive nonlymphoid cells play a role in the development of T cells in the fetal thymus, and that "thymus processed" T cell progenitors (but not the more immature progenitors in the fetal liver or bone marrow) are self-Ia restricted in their differentiation.
Two major classes of mitogen-reactive B lymphocytes defined by life span.
Authors: Freitas A A , Rocha B , Coutinho A A
Published: December 2022

The relative numbers of short- and long-lived mitogen-reactive B cells in the peripheral pool were evaluated by studying the decay of lipopolysaccharide (LPS)-reactive B lymphocytes in LPS nonresponder, histocompatible hosts for periods of up to 3 wk after cell transfer. The results obtained demonstrate the existence of two major classes of mitogen-reactive B cells defined by life span. The "short-lived" cell component comprises about 80 to 90% of the reactive cells and decays with a life expectancy of 18 to 24 hr. The long-lived cell component, with life expectancies of 10 to 20 days, comprises about 10 to 20% of the reactive cells and is preferentially enriched in circulating B cells. The present ratio for short- and long-lived B cells implies a highly dynamic state for the immune system which must be advantageous in the selection of available repertoires.
Immunoglobulin light and heavy chain isotypes in skin diseases: restricted distribution in bullous pemphigoid and linear IgA bullous dermatosis.
Authors: Flotte T J , Baird L G
Published: December 2022

To assess the heterogeneity of immunoglobulins involved in various skin diseases, direct and indirect immunofluorescence studies of skin biopsies and sera, respectively, for kappa and lambda light chains, were performed. The anti-basement membrane zone (anti-BMZ) antibodies of patients with bullous pemphigoid showed a predominance of kappa light chains, and patients with linear IgA bullous dermatosis showed a predominance of one light chain that was sometimes kappa and sometimes lambda. The bullous pemphigoid autoantibodies were then studied for IgG subclass distribution; a predominance of IgG4 was found. Although other explanations are possible, the light chain restriction in bullous pemphigoid most likely reflects heavy chain restriction and preferential association of heavy and light chain isotypes. The basis of the heavy chain restriction is not apparent. The light chain restriction in linear IgA bullous dermatosis may represent a restricted idiotypic repertoire.
Human alveolar macrophages: HLA-DR-positive macrophages that are poor stimulators of a primary mixed leukocyte reaction.
Authors: Lipscomb M F , Lyons C R , Nunez G , Ball E J , Stastny P , Vial W , Lem V , Weissler J , Miller L M
Published: December 2022

Previous studies demonstrated that alveolar macrophages (AM) from most normal human volunteers failed to stimulate the antigen-induced proliferation of peripheral blood T lymphocytes although greater than 90% of AM expressed HLA-DR antigens. The current studies establish that AM also fail to induce allogeneic peripheral blood mononuclear cells to proliferate in a mixed leukocyte reaction (MLR). Suppressive activity by AM was not an explanation for their failure to induce an MLR. Indirect immunofluorescence established the presence of both HLA-DR and DQ antigens on the majority of AM and the persistence of these antigens on cells in culture for up to 6 days, the period of time required to observe a maximal MLR. Metabolic labeling experiments also demonstrated that HLA-DR antigens were synthesized by AM. It was recently reported that AM secrete relatively small amounts of IL 1, an important ancillary signal provided by accessory cells to enhance the stimulation of lymphocyte proliferation. However, addition of optimal concentrations of IL 1 to cultures containing AM failed to enhance the MLR. Thus, there is at least one additional, but as yet undefined, requirement for an accessory cell to induce an optimal MLR besides the display of HLA-D region antigens and the secretion of IL 1. In contrast, AM were effective in specifically stimulating proliferation of alloreactive T cell lines, suggesting that at least some cell lines do not require this nonspecific undefined second signal. We speculate that although AM may not initiate primary immune responses in the lung, they may be important in maintaining immune-mediated inflammatory responses by specifically restimulating already activated T cells.
Ionic requirements and subcellular localization of tubulin tyrosinolation in human polymorphonuclear leukocytes.
Authors: Nath J , Gallin J I
Published: December 2022

We previously reported a specific stimulation of polymorphonuclear leukocyte (PMN) tubulin tyrosinolation as induced by the peptide chemoattractant N-formyl-methionyl-leucyl-phenylalanine (fmet-leu-phe) and the Ca2+ ionophore A23187 that is coupled to the NADPH oxidase-mediated stimulation of the PMN respiratory burst. The present study demonstrates that the presence of extracellular Ca2+ is necessary for fmet-leu-phe- and A23187-induced stimulation of PMN tubulin tyrosinolation, as indicated by the complete inhibition of the response by the addition of 1 mM EGTA to the extracellular medium. Methoxyverapamil (10(-5) M), a putative calcium channel blocker, completely inhibited the fmet-leu-phe-induced stimulation of tubulin tyrosinolation in PMN, but did not inhibit the A23187-induced response. Moreover, the calmodulin-binding drugs, trifluoperazine, fluphenazine, or chlorpromazine, at concentrations of 1 to 10 microM, caused significant inhibition of fmet-leu-phe- or A23187-induced stimulation of tubulin tyrosinolation. In related studies, enzymatic [14C]-tyrosinolation in isolated subcellular fractions of PMN revealed the presence of native tubulin in PMN fractions that were enriched in plasma membranes, the specific granules, or the azurophil granules. Most interestingly, tubulin tyrosine ligase (ligase), primarily a cytoplasmic enzyme, was detected in association with the PMN azurophil granule-rich fraction. Immunoautoradiography with the alpha-tubulin antibody YL 1/2 of isolated PMN subcellular fractions demonstrated a preferential stimulation of tyrosinolation of tubulin associated with the plasma membrane-rich fraction of fmet-leu-phe-stimulated cells. A significant stimulation was also observed in the cytoplasmic tubulin fraction. Consistent with the findings of in vitro tyrosinolation studies with PMN subcellular fractions, tyrosinolated tubulin was detected in the azurophil granule-enriched fractions isolated from both resting and fmet-leu-phe-stimulated cells. The antibody YL 1/2, which reacts with tyrosinolated alpha-tubulin and not with the detyrosinolated form, showed significant cross-reaction with several nontubulin PMN proteins.
Generation of leukotrienes by human monocytes pretreated with cytochalasin B and stimulated with formyl-methionyl-leucyl-phenylalanine.
Authors: Williams J D , Robin J L , Lewis R A , Lee T H , Austen K F
Published: December 2022

The N-formylated tripeptide N-formyl-methionyl-leucyl-phenylalanine (FMLP) initiated the generation of immunoreactive C-6 sulfidopeptide leukotrienes and of leukotriene B4 (LTB4) in a dose-dependent manner from monolayers of human monocytes pretreated for 10 min with 5 micrograms/ml of cytochalasin B. The EC50 for the immunoreactive C-6 sulfidopeptide leukotrienes was 10(-8) M FMLP and for immunoreactive LTB4 was 5 X 10(-8) M FMLP. The maximal response to FMLP occurred within 10 min, and the sum of the two classes of leukotrienes generated was about 1/6 that obtained from monocytes stimulated with calcium ionophore A23187. The requirement for cytochalasin B in order for FMLP, but not the calcium ionophore, to stimulate leukotriene generation is compatible with the ability of cytochalasin B to augment in other cells certain stimulus-specific transmembrane responses that are not dependent on the integrity of the cytoskeleton. Resolution by reverse phase high performance liquid chromatography of the products released from monocytes pretreated with cytochalasin B and stimulated with FMLP or calcium ionophore yielded a single peak of immunoreactive LTB4 eluting at the same retention time as the synthetic standard; immunoreactive C-6 sulfidopeptide leukotrienes eluted at the retention times of leukotriene C4 (LTC4) and leukotriene D4 (LTD4). [3H]LTB4 was not metabolically altered by monocytes pretreated with cytochalasin B and activated with FMLP in comparison with cells treated with buffer alone, whereas [3H]LTC4 was partially converted to [3H]LTD4. The leukotriene-generating response of monolayers of human monocytes pretreated with cytochalasin B to FMLP is receptor-mediated, as indicated by the inactivity of the structural analog N-acetyl-methionyl-leucyl-phenylalanine and by the capacity of the FMLP receptor antagonist carbobenzoxyphenylalanyl-methionine to inhibit the agonist action of FMLP in a dose-response fashion.
Cultured human vascular endothelial cells acquire adhesiveness for neutrophils after stimulation with interleukin 1, endotoxin, and tumor-promoting phorbol diesters.
Authors: Schleimer R P , Rutledge B K
Published: December 2022

Cultured human vascular endothelial cells obtained from umbilical cord veins were observed to acquire adhesive properties for purified neutrophils after exposure to IL 1, endotoxin, and tumor-promoting phorbol diesters. Adhesiveness induced by IL1 and endotoxin had similar kinetics of onset, producing no change after 30 min incubation and reaching optimal change by 4 hr of incubation. The phorbol diester TPA induced changes in adhesiveness more rapidly, with half maximal increase induced by a 15- to 30-min exposure. TPA, but not IL 1 or LPS, induced significant morphologic changes in the endothelial cell monolayer. None of the stimuli decreased endothelial cell viability. All stimuli induced increased adhesiveness at relevant concentrations, i.e., endotoxin, 0.01 to 1 microgram/ml; IL 1, 0.5 to 2 U/ml; and TPA, 1 to 30 ng/ml. Structure activity relationships among phorbol diesters indicate that the response occurs through a typical phorbol diester "receptor." A protein synthesis inhibitor (cycloheximide) and an RNA synthesis inhibitor (actinomycin D) prevented the acquisition of adhesiveness stimulated by IL 1 and endotoxin but not by TPA. In addition, TPA showed a differential temperature sensitivity in inducing adhesiveness in endothelial cells. IL 1 and endotoxin did not produce the effect with a 4-hr incubation at 22 degrees or 4 degrees C, whereas TPA was effective at these lower temperatures. Purified human IL 2 and recombinant-derived interferon-gamma failed to induce adhesiveness in vascular endothelial cells, indicating that this is not a general property of lymphokines. We conclude that endothelium may, under some circumstances, play an active role in producing a leukocyte infiltrate at a local tissue site by acquiring adhesive properties. The production of IL 1 by tissue macrophages, etc., may serve as an important initiator of an inflammatory cell infiltrate. Finally, an action of tumor-promoting phorbol diesters in increasing endothelial cell adhesiveness, combined with their known effects in activating leukocytes, may help explain the extraordinary inflammatory potency of these compounds.
Targeting of drug loaded immunoliposomes to herpes simplex virus infected corneal cells: an effective means of inhibiting virus replication in vitro.
Authors: Norley S G , Huang L , Rouse B T
Published: December 2022

The goal of our studies was to develop liposomes containing antiviral drugs and targeted with antiviral antibody (immunoliposomes) that would be effective at inhibiting replication of herpes simplex virus (HSV) in vitro. To achieve this, a monoclonal antibody to glycoprotein D of HSV was derivatized with palmitic acid and was incorporated into the lamellae of dehydration-rehydration vesicles. The gD containing immunoliposomes were shown to bind specifically to HSV-infected rabbit corneal cells in vitro, whereas control immunoliposomes prepared with a monoclonal antibody of the same class as the anti-gD failed to preferentially bind to virus-infected cells. The gD immunoliposome binding was inhibitable by pretreatment with rabbit anti-HSV serum but not by aggregated normal serum. Thus liposome binding was judged to represent an antigen-antibody reaction not binding to Fc receptors expressed by cells infected with HSV. Immunoliposomes loaded with iododeoxyuridine (IUDR) leaked drug rapidly at 37 degrees C, whereas acyclovir (ACV)-loaded liposomes still contained 48% of drug after 24 hr at 37 degrees C. The ACV-liposomes retained 44% of drug after 14 days at 4 degrees C. The ability of immunoliposomes to inhibit virus replication was compared with that of untargeted and empty liposomes by means of virus yield assays in vitro, Immunoliposomes loaded with either IUDR or ACV inhibited virus replication, although ACV-containing immunoliposomes were the most efficacious. The implications of our in vitro results for the development of immunoliposomes suitable for the treatment of ocular herpes infection are briefly discussed.
Pathogenesis of the delayed phase of Rauscher virus-induced thrombocytopenia.
Authors: Grau G E , Morrow D , Izui S , Lambert P H
Published: December 2022

BALB/c (H-2d) mice infected with Rauscher murine leukemia virus (RMuLV) developed two phases of thrombocytopenia: an acute phase, probably due to direct virus-platelet interactions, and a delayed phase, starting 2 to 3 wk after virus injection, which was associated with the infection of megakaryocytes by RMuLV and with the expression of RMuLV gp70 and p30 antigens on platelet membranes. This study was concerned with the pathogenesis of this second phase of thrombocytopenia. During this period, the number of marrow megakaryocytes was increased. A peripheral platelet destruction was further indicated by reduced platelet life span. It was shown that radiolabeled platelets, either normal or infected, were submitted to a more rapid clearance in infected recipients than in normal recipients. This might be due to the splenomegaly observed in infected recipients. However, the immediate clearance of gp70+ platelets was more accelerated in infected recipients with high titers of serum anti-gp70 antibodies than in infected recipients without detectable serum anti-gp70 antibodies. In addition, the passive transfer of anti-RMuLV serum to normal BALB/c mice induced a rapid and specific clearance of previously injected radiolabeled platelets expressing RMuLV antigens. In H-2d mice, viral gp70 antigen expression on platelets correlated with the development of delayed thrombocytopenia; but H-2k strains of mice, although susceptible to RMuLV and expressing RMuLV-related antigens on their platelets, did not develop any anti-RMuLV antibodies nor any delayed thrombocytopenia. These results suggest that specific clearance of gp70+ platelets in the presence of significant amounts of serum antiviral antibodies and nonspecific hypersplenism play a role in the development of delayed thrombocytopenia in RMuLV-infected mice.
Isolation and characterization of human T cell lines and clones reactive to rabies virus: antigen specificity and production of interferon-gamma.
Authors: Celis E , Miller R W , Wiktor T J , Dietzschold B , Koprowski H
Published: December 2022

By using a preparation of inactivated rabies virus, the blood mononuclear cells from five rabies vaccine recipients were stimulated in vitro in the presence of interleukin 2. T cell lines that displayed significant proliferative responses to whole rabies virus and to preparations of rabies glycoprotein and nucleocapsid were obtained from all the individuals. Other antigens, such as diphtheria and tetanus toxoids, influenza A virus, hepatitis B surface antigen, and serum albumin, failed to induce the proliferation of the T cell lines. One of these rabies-specific T cell lines was found to proliferate in response to rabies antigens only when the antigen-presenting cells expressed homologous HLA-DR antigens. The use of mouse monoclonal antibodies specific for human T cell surface markers revealed that most of the cells of these rabies-reactive lines were of the helper/inducer class of T lymphocytes. Stimulation of the T cell lines with the rabies antigens induced the production of interferon-gamma, a lymphokine with potent antiviral activity. Several T cell clones were isolated from two of these cell lines, and most of them appeared to be specific for the antigenic components of the viral nucleocapsid. Two T cell clones specific for the rabies glycoprotein were also isolated from one of these lymphocyte interleukin 2-dependent lines. Further in vitro studies with rabies-specific T cells could help us to understand in more depth the role of regulatory T cells in the human immune response to rabies virus.
Biology of cloned cytotoxic T lymphocytes specific for lymphocytic choriomeningitis virus. VI. Migration and activity in vivo in acute and persistent infection.
Authors: Byrne J A , Oldstone M B
Published: December 2022

Cloned cytotoxic T lymphocytes (CTL) specific for lymphocytic choriomeningitis virus (LCMV) were adoptively transferred to syngeneic mice acutely or persistently (carrier mice) infected with LCMV. Although infectious virus was cleared from the spleens during acute LCMV infection begun 24 hr earlier and the spleens remained clear of virus for the 4 days of testing, there was no concomitant reduction of viral titers in lymph nodes. In contrast, adoptive transfer of cloned CTL into animals with persistent rather than acute LCMV infection resulted in deaths of syngeneic but not allogeneic recipients. LCMV-immune spleen cells taken 30 to 50 days after a primary immunization and activated by in vitro stimulation before transfer also caused death of syngeneic carrier mice. However, LCMV-immune spleen cell per se provoked no clinical manifestations when transferred but cleared infectious virus and viral nucleic acid sequences from syngeneic carrier mice. The migration of 51Cr-labeled, LCMV-specific, H-2-restricted cloned CTL was assessed in vivo. The circulation of these CTL clearly differed from that of spleen cells freshly isolated from uninfected mice and from non-LCMV-specific CTL clone. Further, the circulatory pattern of LCMV-specific, H-2-restricted, cloned CTL in carrier mice was markedly different than in uninfected animals; only 7% of the injected cells remained in the lungs of uninfected mice 8 hr after injection, whereas 30% had accumulated in the liver. However, 55% of the cells injected into carrier mice still remained in their lungs 8 to 16 hr later. Hence, LCMV-specific, H-2-restricted, cloned CTL have unique trafficking patterns in the presence of LCMV antigens and immune activities in vivo.