摘要:

A method of isolation has. been developed to purify mast cells from human lung tissue. The purification steps are: (1) dispersion of human lung tissue in single‐cell suspensions by enzymatic digestion, (2) partial purification by counterflow centrifugal elutriation, (3) Percoll gradient centrifugation, and (4) enrichment of the mast cells by affinity chromatography using anti‐human IgE‐Sepharose. Enzymatic dispersion yielded 0.6±0.2×106mast cells per gram wet tissue with purities of 3.3±1.0% (mean ±SIMn=3). Elutriation and gradient centrifugation yielded 0.36±0.05×10% mast cells per gram lung tissue in fractions with purities of 30.8±10.7%. Enriched mast cell Fractions were combined, and disposed of contaminating cells by affinity chromatography, thereby yielding 0.25±0.03×106mast cells per gram lung tissue, and improving the purity to 75.3±8.3%. The purified mast cells were intact and vitality exceeded this way from 1 g wet lung tissue 0.25±0.03×106mast cells may be isolated with a mean recovery of 41.7±2.4% and a mean p

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02316.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The effect of polyclonal (anti α and β chain) and monoclonal (anti α chain) antibodies lymphocyte Function‐associated antigen 1 (LFA‐1) on I cell activation was studied. When added at the beginning of activation but not after 24 h or later the antibodies as well as the F(ab')2or Fab fragments of polyclonal antibodies inhibited concanavalin A (Con A)‐induced proliferation, interleukin 2 (IL‐2) production, and the expression of receptors for IL‐2 and transferrin. The inhibitory effect reached a maximum at the same time as optimal proliferation (72 h) Inhibition of proliferation lasted for 5 days or longer, although IL‐2 production was only inhibited during the first 48 h of culture. Receptors for IL‐2 and transferrin were re‐expressed to the original level after 3 days of activation. Addition of external IL‐2 at the beginning of the anti‐LFA‐1 containing culture prevented the inhibition of IL‐2 receptor expression, while inhibition of transferrin receptor expression was unaffected, supporting the conclusion that the‐expression of these two receptors is regulated by partially independent signals. The polyclonal and monclonal anti‐LFA‐1 antibodies also inhibited phorbol ester (PMA)‐dependent OKT3 activation of highly purified T cells. The results suggest that the LFA‐1 antibodies block an early step in the reactions necessary for IL‐2 production, and that the LFA‐1 molecule participates not only in T cell‐accessory cell interaction but also in T‐T interaction durin

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02317.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The regulation of IgE synthesis in vitro and in vivo by schistosomula‐released products (SRP) has been shown to be dependent on the presence of serine proteases. The present paper concerns the characterization of the enzymes involved. The labelling of SRP with [3H]di‐isopropyl phosphofluoridate revealed two molecules, one major with an MW of 27,500 and one minor with an MW of 29,000. The same pattern was obtained by labelling of schistosomula or cercariac surfaces as well as of the total schistosomulum homogenate. The properties of these enzymes were studied by means of various specific substrates or inhibitors of serine proteases. The specificity was relatively narrow, but had some similarity with trypsin. When added to lymphoid rat cells in culture, SRI induced an increased expression of receptors for the Fc fragment of IgE (FcεRII). This suggests that the IgE‐enhancing property of SRP was due to serine protease activity which may act by enhancing the lymphocyte

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02318.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

A case of T cell prolymphocytic leukaemia of helper/inducer T cell phenotype with IgM hypergammaglobulinaemia in a 65‐year‐old Saudi man is presented. The neoplastic T cells in the patient had an OKT3+, OKT4+, OKT8−, OKT11+, OKCLL+, OKIa1+, OKDR+, Tac+phenotype. The presence of OKIa1+, OKDR+, and Tac+antigens indicates that the leukaemic T cells were in an activated state. The cells responded to phytohaemagglutinin, but showed a diminished response to concanavalin A. The reduced interleukin 2 receptor expression and interleukin 2 production were associated with defective concanavalin A‐induced proliferation. There was suppression of mixed lymphocyte culture reaction between the patient and two healthy donors in response to concanavalin A. In vitro immunoglobulin production experiments demonstrate that the leukaemic T cells provided an enhanced helper cell function to produce IgM, and suppressor cell function to produce IgG immunoglobulins by pokeweed mitogen‐induced normal B lymphocyte differentiation. In this study we also found that the patient's T cells proliferate in response to lipopolysaccharides, thus providing the first evidence that leukaemic (OKT4+) T cells from a prolymphocytic leukaemia patient can be stimulated by lipopolysa

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02319.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The lectins fromDatura stramonium, Lycopersicon esculentum, andSolanum tuberosumare structurally related and possess a similar carbohydrate specificity, yet theDaturalectin is mitogenic for human lymphocytes while the other two are not. However, theDaturalectin was found to antagonize blast transformation induced by purified protein derivative (PPD), even within the concentration range at which it was optimally mitogenic on its own. The presence of a submitogenic concentration of the phorbol ester 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA) enormously enhanced the mitogenic activity ofDaturalectin. No such synergy was observed, however, between tomato lectin orS. tuberosumagglutinin (STA) and TPA, nor betweenDaturalectin and the calcium ionophor

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02320.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The analysis of T lymphocytes infiltrating tissues afflicted by autoimmune diseases may provide major clues towards understanding the pathogenesis of such diseases. Currently the best approach to studying heterogeneous populations such as T lymphocytes involves long‐term culture and cloning. In order to grow and clone T lymphocytes, regular restimulation with the specific antigen is essential, otherwise growth will stop and/or specificity may be lost. In autoimmune diseases the antigens involved in triggering the immunological reaction of T cells are usually unknown. Therefore an alternative way of stimulating T lymphocytes without loss of specificity is clearly needed. Here we describe the cloning and expansion of antigen‐specific T cell clones from the blood of a healthy donor to sizeable numbers of cells (>108) by means of anti‐CD3 monoclonal antibody and recombinant IL‐2 The results obtained showed that this approach can be used to clone and ‘expand’ T lymphocytes that retain antigen specificity over a prolonged period, in this case over 10 weeks. This technique has been used to clone and expand T lymphocytes infiltrating the affected tissues in a variety of autoimmune disorders such as Hashimoto's thyroiditis, Graves' disease, and rheumatoid arthritis, and is an efficient method of propagating T cells, by mimicking the antigen

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02321.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Evaluation of lymphokine production by individual activated T cells is necessary to characterize their growth requirements. We have studied interleukin 2 (IL‐2) secretion by mature T lymphocytes using a high resolution assay system with the following characteristics: (a) a threshold of IL‐2 detection 25 times lower than classic IL‐2 titration: (b) the ability to discriminate between IL‐2 and IL‐4 activities: (c) absence of ‘in situ’ IL‐2 absorption; (d) IL‐2 production revealed al the single cell level By this method an average of 10% of spleen cells, and 75% of L3T4 cells were detected as producers in a concanavalin A (Con A)‐dependent T cell activation system. Our results also suggest the complete restriction of II ‐2 secretion to cells with this phenotype. Therefore, factors other than IL‐2 must play a major role in Lyt 2+mitogen‐driven, helper‐indep

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02322.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

We have assessed early changes in quiescent B cells Following cognate and bystander interaction with cloned helper T cells. Variables monitored include Ia expression, blastogenesis, G0to G1transition, and progression through cycle. We have also assessed the antigen specificity. In restriction, and dependence on membrane immunoglobulin cross‐linking of both generation and delivery of effectors that mediate B cell response. The results demonstrate that antigen presentation by quiescent B cells to T cells resulting in the generation of effectors that activate B cells is Ia‐restricted and dependent on antigen and an (mIgM and mIgD) crosslinking signal However, once generated, T cell effector functions act independently of Ia haplotype to promote Ia expression, blastogenesis, and G0to G1, transition by most small B cells. Although these responses can be mediated in T cell supernatants, further progression of B cells through S, G2, and M is only efficient when Thcells are present in cultures. Thus, results suggest that one or more Ia unrestricted, labile and/or cell‐associated factors, not active in most conventional T cell supernatants, are necessary to stimulate proliferation of small B

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02323.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Peripheral blood lymphocytes from patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), progressive systemic sclerosis (PSS). Reiter's disease, osteoarthritis, and from healthy volunteers were investigated for interferon‐γ (IFN‐γ) production after mitogen activation. Phytohaemagglutinin stimulation revealed an impaired IFN‐γ production in RA, SLE, and PSS but normal levels to Reiter's disease and osteoarthritis. In RA this deficiency was also seen after pokeweed mitogen. OKT3, and concanavalin A activation. No major differences were found in interleukin 2 (IL‐2) production and cell proliferation. The IL‐2 receptor expression was reduced on stimulated RA lymphocytes. The deficient IFN‐γ production was compensated in RA by co stimulation of PHA or OKT3 with phorbol myristic acetate (PMA). In addition, the combination of the calcium ionophore A 23187 and PMA induced a strong IFN‐γ secretion in all patient groups a

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02324.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The biosynthesis and secretion of the third component of complement (C3) has been studied with the macrophage cell line J774.2. C3 is initially synthesized as a single polypeptide chain precursor termed pro‐C3, of relative molecular weight (Mr) 170,000 that is post‐translationally modified by proteolytic cleavage into two polypeptides linked by disulphide bonds. The larger polypeptide, termed the alpha chain, has anMrof 110,000–115,000, while the smaller beta chain has anMrof 55,000–60,000. Pulse‐chase experiments indicate that the proteolytic processing of pro‐C3 occurs intracellularly, just prior to secretion. Unlike human C3, which has carbohydrate on both the alpha and beta chains, only the alpha chain of murine C3 is glycosylated. The carboxylic ionophores monensin and nigericin totally inhibit the proteolytic processing of pro‐C3 at a concentration of approximately in 10−6M. This block on proteolytic processing was shown not to be mediated by changes in intracellular pH induced by the disruption of proton gradients. Rather, data from experiments using carboxylic ionophores and other perturbants of cellular physiology indicated that the enzyme(s) responsible for the proteolytic cleavage of pro‐C3 either reside in a cellular compartment with a neutral pH or are proteinases active over a relative

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02325.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY