摘要:

An in vitro lymphocyte‐mediated cytotoxicity assay using [3H]proline‐labeled target cells is described. The assay, modified from an original procedure of Bean et al., assesses the release of [3H]proline by filtering the total culture fluid containing both trypsinized tumor cells and effector cells. Filtration is performed with a semiautomatic harvesting device using low suction pressure and large‐diameter glass filters. Pretreatment of filters with whole serum diminishes adsorption of cell‐free radioactive material considerably and thus increases the sensitivity of the assay. Nearly 100% of the radioactivity could be recovered with this harvesting device. The technique allowed the detection of cytolytic activities of lymphocytes after 6 h of incubation. Lymphocytes from patients with primary malignant melanoma showed a significantly higher cytolytic reactivity(P<0.001) than normal donors’ lymphocytes against three different melanoma cell lines. In a series of parallel experiments on 36 patients and IS normal donors, this modification of the [3H]proline test was compared with three different assays: the conventional microcytotoxicity test of Takasugi and Klein, the original [3H]proline microcytotoxicity test of Bean et al., and the viability count of tu

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00300.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

Experiments were conducted to ascertain to what extent the mitogen response of human peripheral T cells is influenced by contaminated, nonresponding leukocytes, T‐ and B‐lymphocyte preparations, extensively depleted of monocytes, exhibited extremely poor responses to pokeweed mitogen and concanavalin A compared with the original unfractionated cell population. A strongly reduced phytohemagglutinin response was noted in the B‐cell but nut in the T‐cell fraction. Mixtures of cell preparations enriched in T or B cells yielded stimulation higher than expected. Mitomycin‐C‐treated T‐cell preparations mixed with untreated preparations enriched in B cells gave slightly higher responses than expected. However, highly enhanced responses were observed when untreated T cells were mixed with mitomycin‐treated cell suspensions enriched in B cells. A similar enhancing effect was noted when mitomycin‐treated cell suspensions enriched in adherent cells were added. It is concluded that the mitogen reactivity of T cells, on a cell‐for‐cell basis, may be enhanced tenfold or more by cocultivation with unresponsive non‐T leukocytes. A reduced mitogen reactivity of peripheral lymphoid cells from patients with various types of disease may thus either reflect a decreased proportion of responsive lymphocytes or unresponsive non‐T lymphocytes capable of enhancing mi

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00301.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

A competitive inhibition assay was used to define the specificity of the in vitro cytotoxicity of activated lymphocytes. Human and mouse effector cells were produced by prestimulation with phytohemagglutinin, pokeweed mitogen, or purified protein derivative for several days. The addition of increasing numbers of unlabeled Chang cells to a fixed number of stimulated human lymphocytes and51Cr‐labeled Chang cells gradually decreased chromium release. Admixture of unlabeled human bladder tumor cells, lung cells, or monkey kidney cells had a similar effect, whereas mouse L cells were not inhibitory. The reverse was found to be true when stimulated mouse lymphocytes and51Cr‐labeled mouse L cells were incubated. In this case, the addition of unlabeled L or rat tumor cells effectively inhibited51Cr release, whereas human or monkey cells were less inhibitory. Both human and mouse cells inhibited the cytotoxicity of human effector lymphocytes towards mouse L cells. The cytotoxic effects of mouse lymphocytes on human cells were also decreased by addition of unlabeled human, monkey, or mouse cells, but in this case human cells were most inhibitory. The results indicate that activated lymphocytes recognize several surface structures on the target cells. Some of these structures may be shared by cells of different species origin, whereas others seem to be unique for cells from phylogenetically closely related spec

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00302.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

Ox etythrocytes, sensitized with IgM‐type rabbit antibodies, were found to form rosettes with human blood lymphocytes. Granulocytes, monocytes, and macrophages appeared to lack this property. The rosette formation could be inhibited with human IgM and its Fc fragments. IgM‐binding lymphocytes were also present in cell suspensions isolated from human tonsils and thymuses and among blood lymphocytes cultured in vitro. The outcome of combined marker tests and cell separation experiments disclosed that the resetting lymphocytes belong to the T‐lymphocyte popul

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00303.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

A simple technique for organ culture of the thymus of 14‐day‐old mouse embryos is described. It allowed a characteristic time‐dependent development of the immature thymus into a lymphoid thymus with large numbers of nondividing small lymphocytes. Most of these cells carried the T‐lymphocyte antigenic markers Thy‐1 and TL, and all expressed H‐2 antigens as determined in cytotoxicity assays. They probably developed from precursor cells without detectable Thy‐1 and TL. This development appeared to be dependent on the thymic microenvironment, since it also occurred in serum‐free organ cultures and in cultures with medium supplemented with serum

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00304.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

The strong cross‐reaction between rabbit and human β2‐microglobulins, as detected by goat antisera, has permitted the development of heterologous radioimmunoassays to detect homologues of β2‐microglabulin (β2μ) in the sera of various vertebrate species. The assay based on the inhibition of the binding reaction between goat anti‐rabbit β2μ and human β2μ divided the sera examined into two groups, one giving high levels of inhibition (∼80%), and the other lower levels (∼20%). There was no evident correlation between this division and phylogenetic classification. When the same group of sera was tested using the assay based on the inhibition of binding of rabbit β2μ to goat anti‐human β2μ, a wide range of maximal levels of inhibition was observed, with clustering of phylog

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00305.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

A marked prozone effect was observed in indirect immunofluorescence (IFL) with rabbit antisera against rat renal proximal tubular epithelial brush border (BB) antigens: anti‐BB antibodies were not detectable if used in high concentrations but were readily demonstrated if sufficiently diluted. The prozone effect occurred in spite of demonstrated binding of anti‐BB antibodies to target antigens. No prozone was seen when direct IFL was used, and ‘cross‐reaction’ antigens in small‐intestinal epithelial brush border were detectable by indirect IFL without prozone. It was concluded that the anti‐immunoglobulin conjugates were unable to reach antigenic determinants on tightly clustered immunoglobulin molecules. Thus prozone effects seem to depend primarily on the density of antigen determinants in the t

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00306.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

H‐2fais a spontaneous mutation that occurred in an (A.CA × A)F1female mouse and was later transferred on an A‐strain background [congenic line A.CA(M506) or M506]. Skin grafts exchanged between M506 and A.CA strains are rejected within 4 weeks after grafting. Significant reaction is observed when these two strains are used as responders and stimulators in mixed lymphocyte culture or in a splenomegaly graft‐versus‐host assay. The mutant antigen induces strong cell‐mediated lymphocytotoxicity. The mutation maps in theKend of theH‐2complex. The capacity of mutant antigens to induce relatively strong lymphocyte proliferation is explained by hypothesizing that the T‐cell receptors reacting withK‐ orD‐region products can more easily recognize subtle than drastic alloant

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00307.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

Lymph from pigs given bacillus Calmette‐Guérin (BCG) and stimulated in the drainage area of a cannulated lymph node with tuberculin (PPD) had potent in vitro macrophage migration inhibitory activity. A similar observation has previously been made in sheep by other workers. In the sheep, part of, and in the pig. almost all of, this inhibitory activity was produced, not by a factor resembling the so‐called ‘migration inhibitory factor’, but by a mechanism involving a protease and fibrinogen. It is suggested that exudate cells, having potent coagulation‐promoting activity, can promote the deposition of fibrin about themselves, and thus ‘trapped’ are una

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00308.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY

摘要:

Antibodies to human IgG (rheumatoid factors (RF)) from serum Han resembled the ‘intermediate’ complexes of sera from some patients with rheumatoid arthritis. These RF were isolated from heat‐inactivated serum by affinity chromatography on agarose‐coupled human IgG. Indirect immunofluorescence revealed that the isolated RF cross‐reacted with cell nuclei from many species. Virtually all the antinuclear factor (ANF) activity of this serum was extracted by IgG‐agarose. Alternative explanations of this phenomenon, such as nonspecific binding of a separate population of ANF, binding of ANF to the immunosorbent via an IgG RF‐IgG ANF immune complex, or presence of nuclear antigens on the immunosorbent, were ruled out. The cross‐reacting antibodies possessed both k and λ light chains and predominantly γ and some μ heavy chains, indicating that they were of polyclonal origin. The antinuclear activity was present in the F(ab′)2fragments. The interpretation of this strange cross‐reactio

ISSN:0300-9475    

DOI:10.1111/j.1365-3083.1976.tb00309.x

出版商:Blackwell Publishing Ltd    

年代:1976

数据来源: WILEY