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R M Binns

Publications and source records attributed to R M Binns.

At least 91 records · Page 5Linked to original sources
PubMed1982

Subpopulations of pig blood E-rosette-forming lymphocytes and thymus-dependent null cells: separation by nylon wool columns, rosette formation and macrophage-dependent mitogen and antigen responsiveness.

Pig peripheral blood lymphocyte subpopulations were characterized by their adherence to nylon wool. Thus whilst B cells, defined by surface Ig and Fc receptor, were predominantly adherent, thymus-dependent Null cells and the T cells forming rosettes with sheep red blood cells in dextran (DS+) were predominantly non-adherent. There was a direct relationship between the size of the non-adherent fraction and the proportion of the total DS+ cells which were in the non-adherent fraction. Using nylon wool and/or DS rosette separation, lymphocytes from normal and BCG sensitized animals were stimulated in vitro with phytohemagglutinin (PHA), concanavalin A (Con A) and tuberculin (PPD). The cells responding to PHA and Con A (T cell mitogens in other species) were DS+ lymphocytes. Non-rosetting, adherent (B cells) and non-adherent (thymus dependent Null cells) lymphocytes were unresponsive, even in the presence of homologous alveolar macrophages at levels which greatly enhanced the PPD response. Cells responding to PPD were present in the DS+ and DS-, adherent and non-adherent fractions but their response was apparently controlled by macrophage dependence and suppression. Techniques are described for the preparation and storage in liquid nitrogen of alveolar macrophages resulting in high numbers of viable cells, capable of helping in vitro responses, even after 3 years of storage.

Animals↗
PubMed1982

Mouse red blood cell rosettes: human B and some T lymphocytes express receptors for mouse erythrocytes in the presence of Ficoll.

Inclusion of 14% Ficoll in the assay of rosette formation of human lymphocytes with mouse red blood cells (RBC) markedly enhances the cellular interaction. Both the strength of the rosettes (number of attached RBC) and the number of reacting lymphocytes are increased significantly, irrespective of the lymphocyte source--blood, lymph node and tonsil. In the presence of Ficoll, most of the sIg-bearing B lymphocytes [expressing receptors for Fc(IgG) and/or C3] and a significant number of the sheep RBC-rosetting T lymphocytes (particularly the nylon wool non-adherent subset from the tonsil), express receptors for the mouse erythrocyte.

Age Factors↗
PubMed1981

A major subpopulation of Fc receptor-bearing lymphocytes revealed by rosette formation in dextran media: studies with pig, sheep and rat lymphocytes.

Blood lymphocytes from young pigs which formed 9.1 +/-0.7% Fc rosettes (mean +/- S.E., range 4.1-16.0), with rabbit and pig immunoglobulin-coated indicator cells used in optimum conditions in phosphate-buffered saline (PBS), formed 32.6 +/- 1.8% (16.9-44.3) in the presence of 4% dextran (DFc). The proportion of DFc+/Fc+ lymphocytes varied from 2.2 to 5.9 (3.8 +/- 0.3). Compared with the PBS test, in dextran rosettes are formed with more lymphocytes and with red cells coated with less immunoglobulin. Ficoll at 14% gave similar enhancement. Dextran enhancement of Fc rosettes was also observed with sheep PBL, but not with rat thoracic duct lymphocytes. The FC portion of IgG is responsible for both Fc and DFc rosettes. Thus most Fc and DFc rosettes, formed with serum antibody-coated bovine RBC, are revealed by IgG coated but not F(ab')2-coated BRBC, and show dose-dependent inhibition in the presence of free pig IgG (complete at 5-10 mg/nl) but not IgM (up to 2 mg/ml). Separation of lymphocytes on nylon wool suggests that DFc rosettes are formed by B cells and some T cells. Over half of the weak Fc rosette-forming lymphocytes (DFc-Fc) elute in the non-adherent fraction, which contains few B cells, and therefore are a subpopulation of T cells.

Animals↗
PubMed1981

Characterization of lymphocyte subpopulations in sheep by rosette formation, adherence to nylon wool and mitogen responsiveness.

Sheep peripheral blood lymphocytes have been studied using a number of surface markers. Thus 16.6 +/- 2.4% (mean +/- S.E.) were surface immunoglobulin positive (sIg+) by direct immunofluorescence, 35.9 +/- 2.1% formed Fc rosettes with bovine red blood cells (RBC) sensitized with rabbit antibody (Fc+) and 28.4 +/- 2.0% formed rosettes with sheep red blood cells (RBC) in the presence of 4% dextran (DS+). The percentage of both Fc+ and DS+ lymphocytes tended to increase with age of the animals. Demonstration of these markers allowed computation of two further subpopulations: null cells lacking sIg and a receptor for sheep RBC, and Fc.null cells lacking a receptor for Fc and sheep RBC. The former population, which contained a proportion of Fc+ lymphocytes comprised 49.8 +/- 3.8% of blood lymphocytes and the latter 38.4 +/- 3.0%. Separation on nylon wool columns, selective rosette enrichment and depletion on density gradients and stimulation with phytomitogens have shown sIg+ and Fc+ lymphocytes to be nylon wool adherent and unresponsive to phytohaemagglutinin (PHA) and Concanavalin A (Con A) and DS+ lymphocytes to be nylon wool non-adherent and responsive to PHA and Con A. The data also indicates a major overlap of the lymphocyte subpopulations bearing sIg and Fc which are apparently B lymphocytes. Moreover these data support the contention that E-rosette formation with sheep RBC in the presence of dextran is a marker for sheep T cells. The data also indicates tha Fc.null cells are T cells, eluting in the non-adherent fraction from nylon wool. It is probable that a proportion of these cells bear a SRBC receptor too weak for present detection methods.

Animals↗
PubMed1981

Migration of fluoresceinated pig lymphocytes in vivo: technical aspects and use in studies of autologous and homologous cell survival for up to three weeks.

Optimum conditions were determined for FITC and RITC labelling of pig blood lymphocytes with bright fluorescence and high viability and for their enumeration during their long survival after infusion in vivo. For at least 9 days, FITZ-labelled autologous lymphocytes and lymphocytes from MHC-identical donors showed no detectable difference in patterns of survival in the blood. After initial equilibration with extra-vascular lymphoid tissues, the fall-off suggested the expected presence of populations with a variety of life-spans, with some autologous labelled lymphocytes surviving beyond 3 weeks. A trace of cytotoxic antibody specific for labelled cells developed in these pigs later and could be boosted by a further FITC-labelled cell infusion.

Animals↗
PubMed1981

Classification of lymphocytes recirculating in the spleen.

After washing out remaining blood cells using cold perfusate, pig spleens were connected to a closed-circuit normothermic perfusion system. Recirculating lymphocytes migrated out and rapidly established an equilibrium with the splenic extravascular lymphoid tissues. Comparison of the subpopulations among these lymphocytes with those in the blood at time of removal of the spleen, showed the absence of Null cells and reduced numbers of weakly E-rosetting cells among splenic emigrants. The proportion of other B- and T-cell subpopulations were increased in a way which might be explained by non-recirculation of these two subpopulations. Autologous blood lymphocytes labelled with fluorescein isothiocyanate and introduced into spleen perfusions rapidly established equilibrium (approximately 15%-25%, well within 1 hr) and at the end of perfusion were concentrated in the marginal zones. The fluorescent lymphocytes remaining in the perfusate showed a marked enrichment of Null cells suggesting that these cells do not home to splenic extravascular lymphoid tissues in agreement with the hypothesis that Null cells are absent from splenic emigrants because they are nonrecirculating cells.

Animals↗
PubMed1981

In vivo labelling of the spleen and mesenteric lymph nodes with fluorescein isothiocyanate for lymphocyte migration studies.

Lymphocytes in normal young pigs were labelled in vivo with fluorescein isothiocyanate in the spleen using an extracorporeal perfusion system and in mesenteric lymph nodes by direct injection into the nodes. Labelled lymphocytes leave the spleen at a high rate via the splenic vein and migrate to different lymphoid organs. Emigrants from mesenteric lymph nodes left the nodes more slowly and revealed a different homing pattern. Evidence is presented that a considerable number of lymphocytes from the parenchyma leave the nodes via the vein and not by the classical route of recirculating lymphocytes via the efferent lymphatics. Fluorescein labelling of lymphocytes in their normal micro-environment is a suitable method for lymphocyte migration studies.

Animals↗
PubMed1980

Serological and physicochemical reactivity of bovine erythrocytes before and after trypsin treatment.

On partition in aqueous polymer two-phase systems containing dextran, poly(ethyleneglycol) (PEG), and PEG substituted with charged or hydrophobic groups "inagglutinable" ox erythrocytes showed more negative surface charge and less hydrophobicity than "agglutinable" ox erythrocytes. Tryspin treatment of the erythrocytes increased the agglutinability, reduced the negative charge, and seemed to increase the liability to hydrophobic interaction. Haemagglutination of highly negatively charged ox erythrocytes is almost impossible to accomplish by antibody against the ox erythrocytes alone or against IgG antigen linked to the erythrocytes in passive haemagglutination. Likewise, reverse passive (antiglobulin) haemagglutination cannot be accomplished with these cells when immunoglobulin antigens of moderate molecular size (less than or equal to 900,000) are used. However, these cells may be agglutinated when a non-charged carrier such as a bacterium coated with sensitizing antibody (immunoglobulin) is used to bridge the antiglobulin-coupled erythrocytes in a mixed latticer aggllutinate. Such a bridging can also be accomplished by heat aggregation of the immunoglobulin antigen.

Animals↗
PubMed1979

Immunological activity of a T hybrid line. I. Production of an H-2-related suppressor factor with specificity for sheep red blood cells.

T lymphocyte hybrid lines have been produced by fusion of the thymoma BW 5147 with spleen cells of C57BL/10 mice primed to sheep red blood cells (SRBC). The supernatant (culture fluid) of a T hybrid designated A 1 was able to suppress the primary (IgM) and secondary (IgM and IgG) antibody responses to SRBC in vitro. The suppressive activity of supernatants could be titrated to 50% end points at final dilutions of up to 1 : 270. The suppression affected only SRBC and haptens coupled to SRBC, except when used at high concentration when some nonspecific suppression was observed. Absorption of the A 1 supernatant with SRBC removed all activity, while several other species of red cells failed to do so. The suppressor factor present in A 1 supernatant was not removed by anti-Ig adsorbents, but was removed by anti-H-2 antibodies reacting specifically with the haplotype of the spleen cells used in the fusion (H-2b). The cellular target of action of the factor was apparently a B cell, based on absorption with different cell populations. No genetic restrictions in the activity of the factor were found. A 1 cells carried H-2 and Thy-1 alleles of both parental cells and formed rosettes with SRBC.

Absorption↗
PubMed1979

Characterization of pig lymphocyte subpopulations by adherence to nylon wool.

The property of adherence to nylon wool was used to separate and characterize pig blood lymphocyte subpopulations. Thymus-dependent null cells proved the least adherent lymphocytes, and B cells, bearing surface Ig or Fc receptor, the most adherent. sIg+ lymphocytes detected by immunofluorescence and by 51Cr release cytotoxicity showed similar frequency and adherence properties. Lymphocytes rosetting with SRBC in PBS or in dextran had similar adherence patterns and showed both non-adherent and adherent components. Lymphocytes bearing major histocompatibility complex (MHC) Ia-like antigens shown by eosin exclusion or 51Cr release cytotoxicity were enriched in the adherent fraction, but also occurred in significant numbers in the non-adherent fraction. Non-adherent Ia+ cells were predominantly sIg- and probably T cells, whereas the adherent Ia+ group included the sIg+ cells and some sIg- cells.

Animals↗
PubMed1979

Comparison of the direct antiglobulin rosetting reaction (DARR) and direct immunofluorescence (DIF) for demonstration of sIg-bearing lymphocytes in pigs, sheep and cattle.

Tests with untreated and trypsin-treated red cells (rbc) from a variety of species showed that anti-Ig-coupled pig RBC are good indicator cells for the study of ruminant blood sIg + lymphocytes by the DARR test; coupled donkey and rabbit RBC are suitable for investigating pig lymphocytes. The different species showed the following percentages of sIg + lymphocytes (M +/- SE) by direct immunofluorescence (DIF) and the direct antiglobulin rosetting reaction (DARR) respectively:pigs 9.2 +/- 0.7% and 16.3 +/- 1.2%; sheep 20.2 +/- 1.2% and 33.1 +/- 1.6%; Cattle 13.5 +/- 1.4% and 28.9 +/- 3.5%. The mean ratio of sIg + lymphocytes shown by the two tests (DARR/DIF) for each species was 1.80 +/- 0.08 for pigs, 1.73 +/- 0.7 for sheep and 2.15 +/- 0.18 for cattle. Preincubation of pig and sheep lymphocytes at 37 degrees for 1 h did not alter the proportion of sIg + lymphocytes detected by either test. Thus the DARR test reveals a further population of sIg + lymphocytes in addition to that detected by immunofluorescence, whose number is proportional to the B population as measured by DIF and whose sIg is intimately associated with the membrane.

Animals↗
PubMed1978

Sheep erythrocyte rosettes in pigs, sheep, cattle and goats demonstrated in the presence of dextran.

Formation of rosettes with sheep RBC by blood lymphocytes from young pigs was increased from 24.4% +/- 2.2 (mean +/- S.E.) in PBS to 54.1% +/- 1.9 in the presence of dextran. This increase was achieved without inducing appreciable rosette formation with other RBC which do not form rosettes in PBS. Lymphocytes which rosette only in dextran are predominant in pigs between 20 and 160 days old, when the peripheral lymphocyte pool is increasing very rapidly. The use of dextran revealed major populations of blood lymphocytes rosetting with SRBC in adult sheep (30.7% +/- 2.0), adult cattle (37.3% +/- 4.2) and adult goats (13.3% +/- 1.2). Proportions of rosette-forming lymphocytes tended to increase with age. In calf lymphoid tissues the distribution of rosette-forming lymphocytes suggested that these were T cells. In the improvement of rosette formation with SRBC, dextran was more effective than foetal calf serum, papain treatment of the SRBC or combinations of these treatments.

Age Factors↗
PubMed1978

Major transplantation antigens of pig kidney and liver. Comparisons between the whole organs and their parenchymal constituents.

We have previously shown that cultured renal and hepatic parenchymal cells of the pig differ in the expression of major transplantation antigens. Antigens apparently analogous to those of the mouse I region were detectable on kidney and some peripheral blood lymphocytes but not on liver, platelets, or other peripheral blood lymphocytes. The present study confirms these findings for liver parenchymal cells and for a mixture of kidney tubules and glomeruli prepared from fresh, perfused whole organs. However, homogenates prepared at the same time from the perfused whole organs showed no antigenic disparity. Whole liver homogenate contains I region analogues that are barely detectable on the parenchymal cell membrane. Liver parencymal cells may possess such antigens in a form not patnetly expressed, or the antigens may belong to other types of cell resident in the liver which are not removed by extensive perfusion. Similar results were obtained with liver homogenates derived from an orthotopic transplant to a fully allogeneic recipient and resident for 37 days, suggesting that blood lymphocytes persisting in the normal liver after perfusion could not account for these findings.

Animals↗
PubMed1978

Contact sensitivity in the pig. II. Induction by intralymphatic infusion of DNP conjugated cells.

Pigs were skin painted with the contact sensitizing agent DNFB and afferent lymph was collected for the next 24 h. The lymph cells carried a small amount of DNP and were able to sensitize 40% of homologous recipients. Peripheral blood lymphocytes conjugated with DNFB in vitro to the same degree as afferent lymph cells sensitized 80% of autologous and 40% of homologous recipients: lymphocytes conjugated at higher levels sensitized 75% of animals in each group. Lightly conjugated cells were capable of survival and able to respond to stimulation by mitogens and their ability to sensitize autologous recipients was abolished by heat killing. Highly conjugated cells were not capable of survival, their sensitizing ability was not altered by heat killing and they were able to sensitize incompatible recipients. It is suggested that highly conjugated cells sensitize by a different mechanism which depends on the cooperation of non-lymphocytic cells, not easily mobilized from lymphoid tissue.

Animals↗
PubMed1978

Contact sensitivity in the pig. III. Induction by intralymphatic infusion of DNP-conjugated cell membranes and soluble proteins, free DNFB and some small molecular weight derivatives.

DNP-conjugated lymph node cell plasma membranes, thymocyte plasma membranes and red cell ghosts were prepared and tested for their ability to induce contact sensitivity, using pigs as experimental animals. Lymph node cell membranes and red cell ghosts were able to sensitize, provided the dose of DNP was very large, but thymocyte membrane failed to sensitize most pigs. DNP-conjugated lymph proteins coming from the site of application of DNFB were also able to sensitize if large amounts were administered, but free DNFB itself, infused directly into an afferent lymphatic, was much more efficient. Since DNFB can often be detected in lymph folowing skin painting, it may conjugate cells within the node which later contact the recirculating population of lymphocytes and so sensitize the animal by a central mechanism. The best-equipped cells would be the macrophage-like lymph cells which are closely related to the epidermal Langerhans cells and are known to migrate to the paracortex of the node.

Animals↗