摘要:

Anti‐mitochondrial autoantibodies (AMA) from patients with primary biliary cirrhosis (PBC) were analysed for fine specificity by immunoblotting and enzyme‐linked immunosorbent assay (ELISA). Inhibition ELISA showed that complex I (NADH‐ubiquinone reductase) from beef heart mitochondria completely inhibited the binding of AMA to mitochondrial inner membranes (SMP), indicating that the major mitochondrial antigens are located in complex I, Immunoblot analysis of beef heart SMP, complex I and the iron sulphur (IP) subfraction of Complex I revealed several antigens, one of which (75 kDa) reacted with all PBC sera but not with the additional autoimmune sera tested. Resolution of SMP or complex I by two‐dimensional electrophoresis yielded in both preparations a polypeptide of 75 kDa with an isoelectric point of 6.4, which reacted with PBC serum and with rabbit antisera raised against the 75,000 subunit of complex I. In immunoblot experiments, the antigenicity of the 75,000 polypeptide in SMP, complex I, and the IP subfraction is increased by prior reduction of the sample with mercaptoethanol. This suggests a similarity to the PBC‐specific ‘M‐2’antigen, which is also sensitive to sulphur reagents. The data indicate that the 75 kDa polypeptide of complex I is a major mitochondrial antigen binding AMA in PBC sera, and allows us to identify the location and probable function of

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01497.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Purified human amyloid protein A (AA) or serum amyloid protein A (SAA) was incubated with normal human high‐density lipoprotein (HDL). After ultracentrifugation the amount of AA or SAA associated with HDL was measured. It was found that the binding capacity of HDL for SAA was higher than that for AA. Incubation of these in vitro associated HDL‐AA and HDL‐SAA complexes with purified apo AI or apo AII resulted in varying degrees of displace‐ment of the associated AA or SAA from HDL. Under the experimental conditions used, apo AI was able to displace AA from HDL, while apo AII was able to displace both SAA and AA. This indicates that the binding capacity of HDL is different for SAA and AA Mouse acute‐phase HDL was isolated and the native complexes were incubated with human apo AII SAA2, the amyloidogenic SAA variant in mice, was displaced from HDL to a greater extent than SAA1, indicating a lower binding capacity for the amyloidogenic SAA variant for the HDL

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01498.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Spontaneous lymphocyte proliferation (SpP), measured in vitro as the rate of [14C]thymidine incorporation in blood lymphocytes, was investigated in non‐infected postoperative patients, infected postoperative patients, and healthy volunteers, with 72, 24, and 3 h of lymphocyte culture. With 24‐h cultures, infected postoperative patients showed 17‐fold higher SpP than non‐infected postoperative patients (2527±1552 versus 151±77 cpm, mean± SD,P<0.001) and 37‐fold higher SpP than healthy volunteers (P<0.001). Postoperative patients without infection had twice as high SpP as healthy volunteers (P<0.001). Lymphocytes harvested after 24 h of cell culture showed significantly higher SpP than corresponding values at 72 and 3 h, in patients as well as in healthy volunteers (P<0.01). Infected postoperative patients showed a higher SpP than non‐infected patients after only 3 h of cell culture (270±192 versus 48±10 cpm,P<0.001). An inverse correlation was observed between the level of SpP and body temperature in patients with postoperative infection (r=‐0.62,P<0.05). The results indicate that lymphocytes are activated by uncomplicated surgery and particularly by postoperative infection, and that characteristics of SpP are reproducible in short cell‐culture periods, which suggests that in vitro measurements of SpP may be of value in the detection of severe pos

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01499.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

In a previous study we observed that human epidermal cell (EC) suspensions containing HLA‐DR‐expressing keratinocytes showed an amplified T‐cell response to purified protein derivative (PPD). To evaluate further the possible immunological importance of class II transplantation antigens on keratinocytes we have compared the T‐cell response to PPD in the presence of the following stimulator cells: EC suspensions from normal skin, or EC from tuberculin‐reactive skin with or without removal of Langerhans' cells. The proliferation of purified T lymphocytes from peripheral blood in response to PPD in the presence of various concentrations of autologous EC was measured by [3H]thymidine incorporation on day 6. In 3 experiments out of 4 the EC from tuberculin‐reactive skin, containing 28–76% HLA‐DR‐expressing cells as judged by immunocytochemistry (which also revealed fairly numerous HLA‐DQ/ ‐DP‐expressing keratinocytes and a slight increase in CD36‐ and CD4‐ but not CD1‐expressing cells), induced a more pronounced T‐cell response to PPD than did normal EC. This was not the case in the fourth experiment, in which a small number of HLA‐DR‐ (15%) and few if any HLA‐DQ‐/‐DP‐expressing keratinocytes were found. Immunomagnetic removal of CD1‐reactive Langerhans' cells from the tuberculin‐reactive EC suspensions resulted in a reduction of the T‐cell response to PPD, in most cases down to background level (T cells alone + PPD). This study does not support the hypothesis that HLA‐DR‐expressing keratinocy

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01500.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

A monoclonal antibody, TM316, IgM, was raised against the human monocytoid leukaemia cell line THP‐1, and was shown to inhibit polymorphonuclear leucocyte (PMN) chemotaxis. The molecular weight (MW) of the protein on the PMN membrane with which TM316 bound was about 78,000. TM316 inhibited the chemotactic response of human PMN induced by at least three kinds of chemotactic factors (activated serum, N‐formyl‐l‐methionyl‐l‐leucyl‐l‐phenylalanine (FMLP), and a lymphocyte‐derived chemotactic factor (LDCF)) to the same extent. The extent of inhibition of chemotaxis by TM316 was strongly correlated with the quantity of cellular surface antigen recognized by TM316, when a cell sorter was used for analysis. TM316 did not alter the number of Fc or complement receptors of PMN, nor did it affect luminol‐enhanced chemiluminescence(CL), lysosomal enzyme release, adherence, or superoxide anion generation by PMN. TM316 seemed to recognize a common surface antigen which was necessary only for the pr

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01501.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

We produced monoclonal anti‐idiotypic antibody against immunoaffinity purified anti‐U1‐ribonucleoprotein (RNP) antibody from a patient (K.T.), by the cell fusion procedure. The specificity of monoclonal anti‐idiotypic antibody (IgG1, x) was determined by inhibition studies. With the monoclonal anti‐idiotypic antibody, cross‐reactive idiotypes on anti‐U1‐RNP antibodies from unrelated patients with anti‐U1‐RNP antibody was detected in 57% of the samples. The anti‐idiotypic antibody specifically suppressed the in vitro production of anti‐U1‐RNP antibody by lymphocytes from the patient K T., and unrelated patients with a cross‐reactive idiotype, in whom idiotype‐reactive T cells were demonstrated. The results indicate that anti‐idiotypic antibody may modulate the regulation of in vitro

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01502.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Treatment of intact cells in the cold with low concentrations (1 mM) of sodiummetaperiodate (PI) selectively oxidizes the surface‐exposed sialic acid residues to the corresponding aldehydes. Such treated tumour cells show greatly enhanced sensitivity to lysis by fresh human NK cells but not to mouse or rat NK cells. Reduction of the PI‐treated cells with sodium borohydride (NaBH4) reduced their NK sensitivity to that of untreated cells. In target conjugate formation assays PI‐treated tumour cells displayed a higher binding capacity than control cells or PI + NaBH4‐treated cells to both mouse and human effector cells. Neuraminidase treatment of K562 and Molt‐4 increased target susceptibility to human NK cells but not to mouse, whereas the susceptibility of Yac‐1 cells was left unchanged using both human and mouse effector cells. The same pattern of reactivity is shown in the target binding assay. These findings indicate that Subtle molecular changes in the surface‐exposed carbohydrates of target cells might have a fundamental impact on their sensitivity to lysis by NK cells from certain species, and that in cross species effector‐target combinations a higher binding capacity is not sufficient for increased

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01503.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

We have previously shown that two distinct mouse placental fractions (PF) are potent immunomodulators in vivo. A 40 kDa PF induces a marked decrease of plaque forming cell (PFC) responses, while a 60 kDa PF increase them. Both effects are specific for the priming antigen. In the present study, these two PF are assayed on a cell‐mediated response to allogeneic cells, i.e. in a local graft‐versus‐host reaction (LGVHR). Mice were primed with allogeneic cells in the presence of various amounts of the 40 kDa or 60 kDa PF, or liver extract (LE) as control. Six days later, their spleen cells were injected into the footpads of F1recipients Precise dose‐response curves were established and the kinetics of the GVH response were carefully followed. Parallel with the modulation of PFC responses, the 40 kDa PF caused a potent inhibition of the LGVHR, while the 60 kDa PF greatly enhanced it. Both effect were specific for the alloantigens injected with the PF. Furthermore, we showed that these modulations were observed whatever the intensity of the GVH reaction, which varied according to the number of primed spleen cells transferred. This report also demonstrates that these PF Can be greatly enriched by passage over affinity columns made of insolubilized lectins. The 40 kDa PF is retained on and can be eluted from columns of insolubilized concanavalin A (Con A) or wheat germ agglutinin (WGA), which indicates that it is a glycoprotein. Conversely the 60 kDa PF does not bind to any of the above lectins and is probably not a glycoprotein. This biochemical purification step is also a good procedure for obtaining an even cleaner separation of the two fractions from each other. Thus, this paper demonstrates that both PF have important regulatory properties on specific cellular immune re

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01504.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

Rat Kupffer cells stimulated with bacterial lipopolysaccharide (LPS) produced high levels of interleukin 1 (IL‐1), as determined by thymocyte proliferation assay. Indomethacin revealed a dose‐dependent augmentation in IL‐l production, in parallel with a dose‐dependent reduction in prostaglandin E2production by Kupffer cells. The addition of exogenous prostaglandin E2, dibutyryl cAMP, or isoproterenol led to a dose‐dependent suppression of IL‐I production. The supernatant from UPS‐stimulated Kupffer cells also contained factors that Inhibited IL‐1‐induced thymocyte proliferation. Upon gel filtration, two inhibitory peaks, at apparent MW of 27,000 and 6000, were obtained. The latter but not the former fraction also affected Interleukin 2 (IL‐2)‐induced thymocyte proliferation. Increasing amounts of IL‐1 overcame the inhibitory activity derived from the 27,000 MW fraction. These results suggest to us that prostaglandin E2and IL‐1 inhibitor released by Kupffer cells may be involved in negative self‐control in regulating IL

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01505.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY

摘要:

We have previously demonstrated that histamine can inhibit human helper T cells by direct interaction with these cells. It has now been investigated whether histamine inhibits lymphokine production by various subsets of CD4+human T cells separated with the Leu‐8 (p80) and Leu‐18 (anti‐CD45R; p220) monoclonal antibodies (MoAb). Histamine was shown to suppress to a similar extent the production of interleukin 2 (IL‐2) and gamma interferon (IFN‐γ) by Leu 3+, Leu 3+8+, and Leu 3+8−cell subsets. Mitogen‐activated, unseparated Leu 3+and purified Leu 3+8−cells produced maximal amounts of IL‐2 after 24 h and IFN‐γ after 72 h of culture. In contrast, the Leu 3+18+subset produced no IL‐2 after 24 h, and maximal amounts of IL‐2 no sooner than 48 h of culture, and only small amounts of IFN‐γ during the entire culture period of 96 h. Histamine suppressed the production of IL‐2 by both subsets, both when produced early (after 24 h), as in the case of the Leu 3+18−subset, and late (after 48 h of culture), as for the Leu 3+18+subset. The IFN‐γ production by the Leu 3+and Leu 3+18−cells and the marginal production by Leu 3+18+cells were significantly suppressed by histamine. Dual staining with Leu 8 and Leu 18 MoAb demonstrated that the Leu 18−cell compartment included both Leu 8+and Leu 8−cells. It was shown that the inhibitory effect of histamine on the early production of IL‐2 and the major production of IFN‐γ by T helper cells is mediated via action on both the Leu 3+18−8−and the Leu 3+18−8+cells. The inhibitory effect of histamine on the late production o

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1988.tb01506.x

出版商:Blackwell Publishing Ltd    

年代:1988

数据来源: WILEY