scholarly journals Delayed-type hypersensitivity to allogeneic cells in mice. III. Sensitivity to cell-surface antigens coded by the major histocompatibility complex and by other genes.

1979 ◽  
Vol 150 (4) ◽  
pp. 965-976 ◽  
Author(s):  
F I Smith ◽  
J F Miller

DTH could be induced to cell-surface antigens coded by either H-2 or non-H-2 genes. Sensitivity was more readily induced across I region than across K- or D-region differences. The presence of an I-region difference during sensitization did not significantly increase the DTH response to K- or D-region-coded antigens. Macrophage processing appeared to be the major route of sensitization to background antigens. Thus, high levels of sensitivity were achieved equally well using viable or disrupted cells, the response was independent of the H-2 haplotype of the allogeneic cells, and transfer was restricted to the K end of the host H-2 complex. Although sensitization to H-2 antigens was obtained with disrupted cells, transfer of sensitivity against viable cells was unrestricted. This suggests a minor role for macrophage processing in sensitization to H-2 antigens.

Author(s):  
K. Chien ◽  
I.P. Shintaku ◽  
A.F. Sassoon ◽  
R.L. Van de Velde ◽  
R. Heusser

Identification of cellular phenotype by cell surface antigens in conjunction with ultrastructural analysis of cellular morphology can be a useful tool in the study of biologic processes as well as in diagnostic histopathology. In this abstract, we describe a simple pre-embedding, protein A-gold staining method which is designed for cell suspensions combining the handling convenience of slide-mounted cell monolayers and the ability to evaluate specimen staining specificity prior to EM embedding.


1991 ◽  
Vol 113 (4) ◽  
pp. 731-741 ◽  
Author(s):  
S H Hansen ◽  
K Sandvig ◽  
B van Deurs

The transfer of molecules from the cell surface to the early endosomes is mediated by preendosomal vesicles. These vesicles, which have pinched off completely from the plasma membrane but not yet fused with endosomes, form the earliest compartment along the endocytic route. Using a new assay to distinguish between free and cell surface connected vesicle profiles, we have characterized the preedosomal compartment ultrastructurally. Our basic experimental setup was labeling of the entire cell surface at 4 degrees C with Con A-gold, warming of the cells to 37 degrees C to allow endocytosis, followed by replacing incubation medium with fixative, all within either 30 or 60 s. Then the fixed cells were incubated with anti-Con A-HRP to distinguish truly free (gold labeled) endocytic vesicles from surface-connected structures. Finally, analysis of thin (20-30 nm) serial sections and quantification of vesicle diameters were carried out. Based on this approach it is shown that the preendosomal compartment comprises both clathrin-coated and non-coated endocytic vesicles with approximately the same frequency but with distinct diameter distributions, the average noncoated vesicle being smaller (95 nm) than the average coated one (110 nm). In parallel experiments, using an anti-transferrin receptor gold-conjugate as a specific marker for clathrin-dependent endocytosis it is also shown that uncoating of coated vesicles plays only a minor role for the total frequency of noncoated vesicles. Furthermore, after perturbation of clathrin-dependent endocytosis by potassium depletion where uptake of transferrin is blocked, noncoated endocytic vesicles with Con A-gold, but not coated vesicles, exist already after 30 and 60 s. Finally, it is shown that the existence of small, free vesicles in the short-time experiments cannot be ascribed to recycling from the early endosomes.


1993 ◽  
Vol 16 (10) ◽  
pp. 1054-1056
Author(s):  
Dai SASAKI ◽  
Satoshi KOSUNAGO ◽  
Takeshi MIKAMI ◽  
Tatsuji MATSUMOTO ◽  
Masuko SUZUKI

1971 ◽  
Vol 134 (4) ◽  
pp. 857-870 ◽  
Author(s):  
Darcy B. Wilson ◽  
Dianne H. Fox

The proliferative reactivity of lymphocytes from rat donors maintained under germfree or conventional conditions was examined in mixed lymphocyte cultures stimulated with allogeneic and xenogeneic cell surface antigens. The results show (a) that lymphocytes from conventionally maintained rats are less reactive to human, hamster, guinea pig, and mouse cell surface antigens than to the major H alloantigens, and (b) that lymphocytes from germfree rats display no demonstrable reactivity to xenogeneic cells, but are quantitatively normal in their response to allogenic cells. The conclusion drawn from these observations is that the circulating lymphocyte pool of an individual consists of a greater proportion of cells reactive to H alloantigens of other members of the same species than to the xenogeneic cellular antigens of members of other species and that this large number of cells is not generated by a mechanism involving immunization to cross-reactive environmental antigens.


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