Surface Antigen Markers Research Articles

Human T-lymphoblastoid cell lines with high and low abilities to produce interferon-gamma constitutively and their susceptibilities to interferon.

A human T-lymphoblastoid cell line, TCL-Fuj, produces large amounts of interferon (IFN)-gamma constitutively. A variant cell line, 2M, was derived from it. Both cell lines express similar surface antigen markers, but differ in surface morphology. Compared with the parent TCL-Fuj cell line, 2M produced less IFN-gamma constitutively but more in response to IFN inducers. The IFNs produced constitutively and on stimulation with inducers were analyzed by SDS-polyacrylamide gel electrophoresis. In TCL-Fuj cells, the constitutive and induced IFNs consisted of the same molecular species (22K and 39K). In 2M cells, smaller IFNs were produced constitutively (18K and 32K) and induction resulted in a marked increase of 22K molecules. These two cell lines also differed in sensitivity to the antiviral activity of IFN. Other T-lymphoblastoid cell lines, HPB-ALL and TCL-Fuj 4 cells, which did not produce IFN-gamma were permissive for vesicular stomatitis virus (VSV) replication; its growth was markedly suppressed by IFN-gamma and -alpha. TCL-Fuj cells were also permissive for VSV, but were not susceptible to the antiviral effect of the IFNs. In contrast, in 2M cells the multiplication of VSV was restricted; the viral yield was further reduced by the IFNs and increased by treatment with anti-human IFN-gamma serum. Several clonal cell lines derived from TCL-Fuj and 2M cells had characteristics similar to the respective parent cell lines. The growth of both cell lines was not affected by IFN-gamma or by -alpha. The separation of antiviral and anti-proliferative susceptibilities was peculiar to 2M cells unlike other cell lines.

Binding of hepatitis virus particles to immobilised Procion Blue-HB and Cibacron Blue 3GA

Two chemically similar triazine dyes, coupled to Chromatographic gels, have been investigated for their ability to bind and retain hepatitis B surface antigen marker of the hepatitis B virus. Both dyes, Procion Blue-HB and Cibacron Blue 3GA, were effective in removing viral particles from plasma protein solutions. A number of combinations of dye and gel support have been tried, to obtain optimal binding. A Cibacron Blue-Trisacryl adsorbent achieved almost complete retention (> 99.5%) of HBsAg before breakthrough occurred. Binding was better at reduced flow rates and on large pore gel matrices, suggesting that steric restrictions limit the interaction. The interaction is directly between immobilised dye and the viral particle, rather than through plasma albumin, which also binds to these affinity adsorbents. These findings are important in the use of immobilised triazine dyes for plasma protein purification. Furthermore, the interaction may be utilised either for the removal of HB viral particles from plasma or as a rapid method of their purification.

The reproducibility of acute lymphoblastic leukemia phenotype determinations: evaluation of monoclonal antibody and conventional hematopoietic markers

Peripheral blood and/or bone marrow lymphoblasts from 25 patients with acute lymphoblastic leukemia (ALL) were tested with a panel of monoclonal antibodies (MoAbs) and conventional hematopoietic markers by three different laboratories. The results were analysed to evaluate the reproducibility of ALL phenotype determinations. Specimens were transported between laboratories by 24-h courier service and were classified on the basis of indirect immunofluorescence MoAb reactivities as follows: B-lineage ALL (BA-1 +T −MCS-2 −); T-lineage ALL (T +BA-1 −MCS-2 −); myeloid antigen ALL (MCS-2 +BA-1 −CALLA −T −) and unclassified ALL (BA-1 −MCS-2 −CALLA −T −). Conventional marker studies for surface immunoglobulin (sIg), cytoplasmic immunoglobulin (cIg), sheep erythrocyte rosette formation (E) and nuclear terminal deoxynucleotidyl transferase (TdT) were also performed. In the cases with sufficient marker data to permit classification, 90% (18/20) were identically classified by different laboratories and this concordance was statistically significant ( p < 0.05). The agreement between laboratories for individual MoAb and conventional marker analyses was statistically significant ( p < 0.05) for all markers with the exception of BA-2, cIg and TdT determinations. Six of 7 discordant BA-2 cases represented BA-2 + evaluations which had subsequent BA-2 − results following specimen transportation. These findings suggest instability of the BA-2 antigen to transport conditions. A similar pattern of positive to negative evaluations following transportation was observed in 5/5 discordant results involving other MoAbs. Disagreement between laboratories for cIg and TdT determinations implies that the detection of cytoplasmic or nuclear antigens may be more prone to subjective interpretation than cell surface antigen marker analyses. Our findings suggest that immunofluorescence marker studies employing MoAbs to cell surface antigens are in general highly reproducible. Our results also indicate that specimen storage conditions and the cellular location of target antigens are important variables which may affect the reproducibility of ALL phenotype determinations.

Transformed lymphocytes from Abelson-diseased mice express levels of a B lineage transformation-associated antigen elevated from that found on normal lymphocytes.

Animals injected with Abelson murine leukemia virus (A-MuLV) rapidly develop fatal bone marrow-derived lymphosarcomas. In all such diseased animals tested, a subpopulation of bone marrow cells expressed a monoclonal antibody-defined, B lineage transformation-associated antigen (6C3 Ag) at levels increased from that detected on normal lymphocytes. Cells bearing a high level of this antigen were found to be transformed as measured by clonal growth in agar, and they expressed surface antigen markers characteristic of early pre-B cells. High-level antigen-expressing cells were found in the bone marrow, lymph nodes, and spleen, but never in the thymus of diseased animals. This distribution agrees with the published pathology of Abelson disease.

Expression of the Chediak-Higashi lysosomal abnormality in human peripheral blood lymphocyte subpopulations

Fusion of lysosomes to form a giant cytoplasmic inclusion is a major abnormality expressed by multiple hematopoietic and non-hematopoietic cell types in Chediak-Higashi (C-H) patients. In this study, the extent of involvement of lymphoid cell subpopulations was defined. Purified populations of B cells, natural killer (NK) cells, and helper T cells were obtained from two C-H patients and normal controls by immunofluorescence staining of their blood mononuclear cells with the monoclonal antibodies HB-2, Leu-7, or Leu-3 followed by fluorescence- activated cell sorting. Cytochemical and ultrastructural analyses as well as functional assays were performed to determine whether or not the C-H lysosomal abnormality was expressed in the different lymphocyte subpopulations. B cells expressed the C-H defect following activation and differentiation. All of the Leu-7+ cells and a significant proportion of the Leu-3+ cells displayed the C-H abnormality. These Leu- 3+ cells share the NK lineage characteristics of granular lymphocyte morphology and the capacity to bind to NK cell targets. In contrast, the C-H abnormality was not observed in non-NK target-binding cells with T helper phenotype, in which clusters of lysosomes formed a normal Gall body. Moreover, T cell functions were unimpaired in C-H patients. These observations raise the issue of the lineal relationship between granular and nongranular lymphocytes typed as T cells on the basis of cell surface antigen markers.

Expression of the Chediak-Higashi Lysosomal Abnormality in Human Peripheral Blood Lymphocyte Subpopulations

Fusion of lysosomes to form a giant cytoplasmic inclusion is a major abnormality expressed by multiple hematopoietic and non-hematopoietic cell types in Chediak-Higashi (C-H) patients. In this study, the extent of involvement of lymphoid cell subpopulations was defined. Purified populations of B cells, natural killer (NK) cells, and helper T cells were obtained from two C-H patients and normal controls by immunofluorescence staining of their blood mononuclear cells with the monoclonal antibodies HB-2, Leu-7, or Leu-3 followed by fluorescence-activated cell sorting. Cytochemical and ultrastructural analyses as well as functional assays were performed to determine whether or not the C-H lysosomal abnormality was expressed in the different lymphocyte subpopulations. B cells expressed the C-H defect following activation and differentiation. All of the Leu-7+ cells and a significant proportion of the Leu-3+ cells displayed the C-H abnormality. These Leu-3+ cells share the NK lineage characteristics of granular lymphocyte morphology and the capacity to bind to NK cell targets. In contrast, the C-H abnormality was not observed in non-NK target-binding cells with T helper phenotype, in which clusters of lysosomes formed a normal Gall body. Moreover, T cell functions were unimpaired in C-H patients. These observations raise the issue of the lineal relationship between granular and nongranular lymphocytes typed as T cells on the basis of cell surface antigen markers.

Virus-Associated Cancers in Greenland: Frequent Hepatitis B Virus Infection But Low Primary Hepatocellular Carcinoma Incidence&lt;xref ref-type="fn" rid="FN2"&gt;2&lt;/xref&gt;&lt;xref ref-type="fn" rid="FN3"&gt;3&lt;/xref&gt;

The incidence of primary hepatocellular carcinoma (PHC) in Greenland between 1960 and 1981 was determined and compared with the rate of this disease in Denmark. The annual age and sex rate (per 100,000) was not significantly different (overall, 1.9 vs. 2.2) despite a large difference in the prevalence of hepatitis B surface antigen (HBsAg) and anti-HBs markers of hepatitis. On the basis of a recent report of a very strong risk of PHC among male HBsAg carriers, 4.0 cases of PHC per year were expected in male Eskimos, but only 0.2 cases per year were observed. The incidence rates of other cancers suggested to be virally associated, including nasopharyngeal carcinoma, salivary gland cancer, and carcinoma of the cervix, were all high in Greenland compared to rates for the Danish population, and these high incidence rates are in accord with the markedly higher prevalences in Greenland Eskimos of viruses with which these other cancers have been associated. Thus Greenland Eskimos do not have a high incidence of PHC despite a high prevalence of HBsAg carriers, which suggests that other carcinogenic factors in this environment may be absent or that protective factors are present.

Natural killer cells are present in the normal human lung but are functionally impotent.

The lung is affected by disorders in which natural killer (NK) cells are thought to play an important defensive role. This study, however, demonstrated that normal lung lymphocytes actually express very little NK cell activity (P less than 0.001 compared with blood lymphocytes). This was true independent of the NK-sensitive target used (K562, U937, MOLT-3, or Daudi). This lack of lung lymphocyte NK activity occurred even though the proportions of lymphocytes in the normal lower respiratory tract with the morphology (large granular lymphocytes) and surface antigen markers of NK cells were similar to that of blood (P greater than 0.5). Although normal lung lymphocytes bound to known NK-sensitive targets, they did not lyse these cells (P less than 0.001 compared with blood), which suggested that the lack of lung NK cell activity resulted from a relative inability of lung NK cells to destroy their targets. While the mechanisms of this functional impotence of lung NK cells are not clear, normal human alveolar macrophages and lower respiratory tract epithelial lining fluid exerted a profound suppressive effect on blood NK cell activity (P less than 0.001 for both) by inhibiting their ability to lyse target cells after binding (P less than 0.001). Though impotent initially, when incubated for 24 h in medium alone, normal lung lymphocytes demonstrated markedly enhanced NK activity (P less than 0.02), which suggested that lung NK cells do have the potential to express NK activity. Interleukin-2 (IL-2) further augmented this effect (P less than 0.05), but gamma interferon did not (P greater than 0.2). Consistent with this observation, lung lymphocytes from patients with active sarcoidosis, a disease in which lung lymphocytes are spontaneously releasing IL-2, did express NK cell activity (P less than 0.01). These studies suggest that although NK cells are present in the normal lung, they are functionally inactive, due, at least in part, to local inhibitory influences. In the presence of IL-2, however, lung NK cell activity is expressed, which suggests that lung NK cell activity can be modulated.

14] Separation of lymphoid cells using immunoadsorbent affinity chromatography

This chapter explores that a key to the understanding of the immune system is the ability to separate and purify lymphoid cells so as to study their characteristics, functions, and interactions. It discusses that a major advance in achieving this aim has been the development of techniques which take advantage of the discriminatory power of antibodies to select for cells bearing a particular type of surface antigen. In many cases this results in the separation of functional lymphocyte subsets since these are usually characterized by particular surface antigen markers. For example, anti-IgM antibodies can be used to separate B cells, most of which carry surface immunoglobulin molecules, from T cells, which do not carry surface immunoglobulin molecules. This chapter describes the use of immunoadsorbent techniques to separate lymphoid cells. In these methods antibodies specific for a lymphocyte surface component are bonded to solid phase matrices to provide a selective anchor for isolating given subpopulations.

Experimental gingivitis in humans. A histochemical and immunological characterization of the lymphoid cell subpopulations.

Cell types in a human experimental gingivitis model were analyzed sequentially on days 0, 4, 8, and 21 of a no oral hygiene period. The cells were characterized using enzyme and surface antigen markers. In all but two of the day 0 specimens a small inflammatory infiltrate was localized immediately beneath the junctional epithelium. These, essentially lymphocytic lesions, consisted of over 70% T‐cells as suggested by the phenotype T‐enzyme +ve/T‐cell surface antigen +ve/B‐cell surface antigen −ve/HLA‐DR −ve/B‐cell subset antigen −ve. At days 4, 8, and 21, although the size of the infiltrate increased, its essential nature did not change. At all times the majority of lymphocytes (over 70%) had the characteristic T‐cell phenotype. These results show that in the developing gingival lesion in humans a T‐cell dominated lesion occurs and persists at least for the 3 week experimental period used in the present study.

Murine T-cell hybridomas that produce lymphokine with macrophage-activating factor activity as a constitutive product

We fused concanavalin A-stimulated spleen lymphocytes of C57BL/6N mice with the AKR T-cell lymphoma BW-5147 G1.4. Four hybridomas expressing both Thy-1.1 and Thy-1.2 surface antigen markers characteristic of both parent types were evaluated for the production of various lymphokines. The resulting T-cell hybridoma constitutively produced a lymphokine that activated murine macrophages to the tumoricidal state without the concomitant production of migration inhibition factor, colony-stimulating factor, or interferon.

Plasmid and surface antigen markers of endemic and epidemic Legionella pneumophila strains.

Environmental and clinical isolates of Legionella pneumophila obtained from the Pittsburgh Veterans Administration Medical Center were studied for the presence of plasmids and for unique surface antigens. The majority of environmental isolates contained a single 80-megadalton plasmid. After an epidemic of nosocomial Legionnaires disease subsided in the Spring of 1981, plasmid-bearing environmental isolates persisted in the environment. Whereas L. pneumophila could not be reisolated from most sites with plasmidless isolates. During this epidemic the attack rate was highest on wards with plasmidless isolates. All clinical isolates were plasmidless. Strains were serotyped by the indirect immunofluorescence method with serum from a single immunized rat which was used both without absorption and after absorption with various plasmid-bearing and plasmidless isolates. These studies suggested that a plasmid-associated surface antigen was present and that the most common plasmidless environmental serotype was similar to the epidemic clinical serotype.

Carrier Status in Hepatitis B: A Modern Scourge With Hope

Viral hepatitis, type B (HBV) is characterized in the acute phase by the appearance of the hepatitis B surface antigen (HBsAg) marker in the serum. Approximately 7% of clinically diagnosed HBV cases continue to demonstrate HBsAg in the serum for months after the clinical symptoms have disappeared. These patients are identified as of hepatitis B. The prevalence of HBsAg carriers in the general population of the United States is estimated to be 0.2% to 0.5%. The majority of these patients are entirely asymptomatic, yet the carrier at times represents a clear health hazard to his contacts. In addition, it is now clear that the HBsAg carrier is at clear risk of liver disease and hepatocellular cancer. There is general agreement at the time of this printing that there is no established way to treat or remove the carrier status in all affected persons. The HBsAg carrier is an important

Preparation of hepatitis B polypeptide micelles from human carrier plasma

Water-soluble protein micelles consisting of the 28,000 (gp28) and 23,000 (p23) molecular weight polypeptide complex of hepatitis B surface antigen can be readily prepared from human plasma containing the surface antigen and other markers of infection with hepatitis B virus. The antigenic activity of the polypeptides was preserved throughout the process of solubilisation and reassociation into micelles. Such preparations are therefore eminently suitable as ‘second generation’ hepatitis B vaccines.

T cell ontogeny. Organ location of maturing populations as defined by surface antigen markers is similar in neonates and adults.

Earlier studies have suggested that splenic T cell populations in nursling mice (less than 18 d of age) have Lyt cell surface antigens that identify them as less mature than their adult counterparts. Studies presented here, however, demonstrate that the expression of the Thy-1, Lyt-1, Lyt-2, and Lyt-3 T cell antigens is virtually identical in 14-d-old and adult T cell populations even though at 14 d, T cells constitute less than 10% of the total spleen cell population. Because the expression of these antigens on the immature (cortical) thymocyte population differs substantially from their expression on peripheral T cells, the maturity of splenic T cells as judged by these criteria is similar in nurslings and adults. Very few cells in the neonatal thymus 4 h after birth correspond, in terms of antigen expression, to the more mature (medullary) thymocyte population of adults, but such cells develop rapidly during the first few days of life. They are present, therefore, sufficiently early to serve as the immediate source of peripheral T cells, as they apparently do in the adult. This then suggests that the locations for the major T cell maturational events are established within the first 2 wk of life of the mouse and maintained as such thereafter. The use of monoclonal antibodies and quantitative immunofluorescence analysis in our studies probably explains the differences between our findings and those reported previously, which relied on cytotoxic depletion by alloantisera and complement to estimate the frequencies of cells carrying the Lyt differentiation antigens in nurslings.

T cell ontogeny: Organ location of maturing populations as defined by surface antigen markers is similar in neonates and adults

T cell ontogeny: Organ location of maturing populations as defined by surface antigen markers is similar in neonates and adults

Plasma membrane enzymes in BALB/c lymphomas with either T or B cell properties, I. 5'-Nucleotidase.

Comparison of membrane bound 5'-nucleotidase activity has been made in crude extracts and plasma membrane fractions from Abelson virus transformed lymphomas, IgM, IgG and IgA producing plasmacytomas and from thymomas with different surface antigen markers. 5'Nucleotidase activity was characterised by the following criteria : 1) optimal pH for enzyme activity, 2) specificity of 5'AMP as a substrate at the optimal pH, 3) specific inhibition of the enzyme by alpha beta methylene ADP, 4) inhibition by EDTA. 5'-Nucleotidase was found to be present on the following B cells : Abelson virus transformed lymphomas, plasmacytomas and LPS-stimulated blasts from nu/nu spleens. The enzyme was also found in one thymoma Lut 13, but it was absent from all the other thymomas studied. A possible relationship between 5'-Nucleotidase and purine metabolism in lymphocyte subpopulations is suggested by the results.

HLA Antigen in Chinese Patients With Hepatocellular Carcinonas&lt;xref ref-type="fn" rid="FN2"&gt;2&lt;/xref&gt;&lt;xref ref-type="fn" rid="FN3"&gt;3&lt;/xref&gt;

One hundred and fourteen Chinese patients with hepatocellular carcinomas (HCC) were studied for alpha-fetoprotein (AFP), hepatitis B surface antigen (HBsAg), anti-HBsAg, and HLA markers. Younger patients with HCC (less than 60 yr old) were more significantly associated with elevated serum AFP (P less than 0.0001) and serum HBsAg (P less than 0.0001) than were older (greater than or equal to 60 yr old) patients. A strong correlation existed between serum AFP and HBsAg among the patients (P less than 0.0001). Of 27 AFP-negative patients, 15 (55.6%) had HLA-B15 compared to 53 of 238 (22.3%) healthy controls (corrected P less than 0.003, relative risk = 4.4). The frequency of HLA-B15 was even higher in the AFP-negative plus HBsAg-negative subgroup (61%). AFP-negative patients also had a significant lack of blood group A.

A surface antigen marker for human monocytes.

Antisera have been raised in rabbits aginst human peritoneal macrophages. After absorption with tonsil cells the sera reacted, by direct and indirect immunfluorescence, with phagocytic mononuclear cells from a variety of tissues and, in addition, stained a small population of nonphagocytic non-T, non-B mononuclear cells present in blood, spleen, and marrow but absent or very rare in tonsil and thymus. The antisera may define a human monocyte-macrophage cell surface differentiation antigen (HuMA) and can be used to deplete or enrich reactive cell population.