ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02331.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Tumour necrosis factor (TNF) or cachectin it an important mediator of endotoxic activity. To investigate the production of TNF from human mononuclear cells (MNC) in response to lipopolysaccharide (LPS), we developed a sensitive and specific enzyme immunoassay (ELISA) and a cytotoxicity bioassay for TNF. The ELISA utilizes the biotin/avidin system and includes four incubation steps. The detection limit was 25 pg recombinant TNF(rTNF)/100μl. There was no interference of medium, serum, plasma, spinal Fluid, or urine and no cross‐reaction with natural or recombinants IL‐1‐α, IL‐1‐β, IL‐2, IFN‐γ, or lymphotoxin (TNF‐β). Recovery of TNF added to the media was 83 123% (n=22). The relative standard deviation within and between assays were 7% and 8% respectively. TNF‐induced cytotoxicity was measured on actinomycin‐D‐treated L‐M mouse fibroblasts. The detection limit in this bioassay was 0.5 U/30μl or 12.5 pg/30μl of rTNF, IL‐1‐α and IL‐1‐β slightly inhibited the cytotoxic activity of rTNF. In this bioassay, cytotoxic activity (50–300 U/ml) was detected only when MNC were stimulated with high concentrations of LPS (100–1000 ng/ml). In contrast, using 0.01–100 ng/ml of LPS, the ELISA detected TNF in a dose‐dependent manner (0.25 ng/ml to 40 ng/ml). It is concluded that TNF is liberated from human blood MNC if stimulated with minute amounts of LPS. It is suggested that human TNF may be secreted in a relatively inactive form or that inhibitors of TNF are generated along with the monokine Because of this, and because commonly used bioassays for TNF fail to distinguish between TNF and lymphotoxin, specific E

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02332.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

A literature search reveals a lack of any previous attempt to Implant foreign tissue into the epididymis. Instead, related studies have been carried out within the adjacent testis. Theoretically, there are peculiar immunobiological factors that can require separate consideration of the two adjacent sites. With this background, we designed the present work by removing the adjacent testis and subsequently used biochemical, histological, and electron microscopic methods to assess the fate of parathyroid allografts buried within the epididymis. Since the concept of immunological privilege is usually explained on the basis of the nature of lymphatic drainage, we also studied the pattern of lymphatic drainage at each allograft bed. In the analysis of our results, the animals were grouped according to the nature of lymph pattern from each transplant bed. We observed that prolonged survival of the tissue allograft occurred only when the lymphatic trunk draining the graft bed bypassed all retroperitoneal nodes and opened directly into the blood stream via the cisterna chyli. This anomalous survival is favoured by pretransplant splenectomy. These results demonstrate that the rat epididymis can sometimes be immunologically privileged.

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02333.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The aging effect on the quantity of intestinal IgA and IgM class‐specific immunoglobulins secreted and the quality of intraluminal intestinal IgA was studied in 5‐to 6‐month old and 24‐to 28‐month‐old male BALB/c mice. The level of IgA increased in the intestinal juice from the aged mice, while the IgM level remained unchanged. These alterations were similar to those found in serum, but the effect of age on serum IgA was profound, almost a threefold increase, in found in serum, but the effect of age on serum IgA was profound, almost a threefold increase, in contrast, to an increase of just under 35% in intestinal secretions. The ratio of the dimeric to total IgA in the small intestine decreased in the older mice, though that of serum was unchanged. In contrast, the total amounts of dimeric IgA in the small intestinal lavage fluid did not change between the two age groups, while the dimeric IgA in serum in older mice were 4.5 times as high as those of young mice. The binding of purified intestinal dimeric IgA to antigens from normal habitant enteric bacteria (γ streptococci andEnterobacter agglomerans) declined in the old mice. In the immunochemical studies, using SDS‐PAGE and isoelectric focusing, the purified intestinal IgA from the young and old mice showed no major difference. Thus, the findings in aged small intestinal perfusates that the increased content of intestinal IgA is due to an increased monomeric IgA, but not to a reduced dimeric IgA, which remains unchanged, and that the binding capacity of the dimeric IgA to the bacterial antigens is diminished, suggest that the level of the matural secretory IgA antibody is decreased. These altered quantitative and functional features of intraluminal intestinal IgA observed in the aged mice appear to be due to the complex heterogeneous effects of senescence on gut‐associated lymphoid tissues, and may contribute to age‐related impairment in gut humor

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02334.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

A filter immuno‐plaque assay was developed which detects alpha interferon (IFN‐α) secreting human peripheral blood leucocytes (PBL) Polyclonal anti‐IFN‐α antibodies were fixed to the nitrocellulose membrane bottoms of 96‐well Millititer plates, which also contained monolayers of glutaraldehyde fixed human WISH amnion cells infected by Herpes simplex virus type 1 (HSV). Such cell are potent IFN inducers and during a 16 h cocultivation with PBL, IFN‐α was absorbed by the membrane‐bound polyclonal antibodies around IFN‐α‐secreting cells. This IFN‐α was detected with murine monoclonal antibodies against IFN‐α and peroxidase labelled antibodies against murine immunoglobulin, using diaminobenzidine as substrate Distinctly stained plaques were seen, the frequency of which gave a minimal estimate of approximately 10 IFN‐α‐producing cells in 104PBL (range in 12 blood donors 2.95–25.1). Fewer plaques than expected were seen at low PBL numbers per culture, one explanation being that cell interactions then limit the IFN‐α response. The immuno‐plaque assay should be useful in further studies of t

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02335.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

In a rat heart transplantation model, permanent graft survival was achieved by anti‐thymocyte globulin (ATG) treatment of Wistar Kyoto (WKy) recipients before grafting with PVG/c hearts. Nine hearts with palpable and electrocardiographic (ECG) function 8–20 months after transplantation were studied with histological and immunohistochemical methods. The dominating findings were fibrosis, intimal thickening, thrombosis of vessels, cellular infiltrates, and a high number of cells expressing class II antigens. These findings suggest that reactions compatible with a chronic rejection can be found in long‐term surviving rat heart allografts. This rat model may thus be used in further studies of a chronic rejection pr

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02336.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The modulation of E receptors on T cells by anti‐CD4 and/or anti‐CD8 is reported. The percentage of E rosette‐forming cells (RFC) was decreased when sheep erythrocyte and mononuclear cells were incubated in the presence of monoclonal antibodies anti‐CD4 and/or anti‐CD8. This inhibition was not due to shedding of the E receptor and it was reversed by 8‐Br.‐3′,5’cyclic guanosine monophosphate (cGMP). In contrast, anti‐HLA antibodies induced an enhancement of sheep erythrocyte receptor expressi

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02337.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Twenty‐one patients treated for active tuberculosis were examined for immune reactivity to purified protein derivative (PPD) and to a purified 32‐kDa protein antigen (P32) fromMycobacterium bovis, strain BCG. Lymphoproliferation of peripheral blood leucocytes to PPD and P32 was positive in 95% and 71% of the patients respectively. A positive IFN‐γ response was detected in 62% against PPD and in 48% against P32. Low blastogenesis and IFN‐γ production were observed, especially in patients with poor general health and advanced tuberculous lesions. Twelve out of twelve (100%) of the tuberculin‐positive healthy volunteers responded to PPD and P32 with mean lymphoproliferation and IFN‐γ values that were higher than in the patient group. Twelve tuberculin‐negative control subjects were completely unreactive to PPD and P32 antigen. On the other hand, IgG antibodies in the serum were detected in 95% of the patients against PPD, in 77% of the patients against P32 but in none of the tuberculin‐positive or negative healthy volunteers. The highest IgG levels against PPD were found in those patients with the lowest in vitro lymphoproliferation and IFN‐γ production (r=‐0.54;P<0.05). Nonspecific interferon production following induction with Newcastle disease virus,Corynebacterium parvum, or phytohaemagglutinin was comparable in the control and patient groups. Finally, low IFN‐α titres were detected in the serum of

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02338.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

The primary structure of rat β2‐microglobulin (β2m) was determined. It is a polypeptide of 99 amino acids with the following sequence: IQKTPQIQVY SRHPPENGKP NFLNCYVSQF HPPQIEIELL KNGKKIPNIE MSDLSFSKDW SFYILAHTEF TPTETDVYAC RVKHVTLKEP KTVTWDRDM. The primary structure was determined by NH2‐terminal sequence analysis together with sequence determination of one cyanogen bromide fragment and one tryptic peptide. Of other known β2m sequences, rat β2m is most homologous to mouse β2m (83% identity). The rabbit, human, and guinea pig sequences are more distant, with 24,27, and 31% differences, resp

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02339.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Polyclonal protein HC‐IgA complexes (HC‐IgA) were isolated from two different serum pools. Their hydrodynamic volumes were found to be slightly greater than that of monomeric IgA but less than that of dimeric IgA. Sodium dodecyl sulphate (SDS)‐polyacrylamide gel electrophoresis of reduced and carboxymethylated complexes followed by immunoblotting showed that the complexes contained normal light and heavy Ig chains, and one polypeptide chain withMt= 90,000, which carried both IgA α chain and protein HC epitopes. Enzyme‐linked immunosorbent assays (ELISA) demonstrated that the isolated HC‐IgA carried about 0.1 and 4%, respectively, of the antibody activities against one carbohydrate antigen (Yersinia enterocoliticaserotype 0:3 lipopolysaccharide) and one protein antigen (rabbit IgG, i.e. antigen for rheumatoid factors) of the IgA populations of the two serum pools. HC‐IgA with rheumatoid factor activity could also be demonstrated in the unfractionated serum pool. The binding of HC‐IgA in the ELISA was not mediated through its protein HC part. The present observations show that HC‐IgA carries antibody activities and constitutes a unique class of IgA complexes, since it does not dissociate under denaturating conditions alter reduction. It may represent a further biological potential of the humor

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb02340.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY