a. 自己抗体産生細胞の同定法

抄録

The mechanism of the autoimmune diseases remains still unknown. It is doubtless, however, that the production of auto-antibody by antibody producing cells is the direct cause of such diseases. The authors intended to identify the auto-antibody producing cell by observing the antigen-antibody reaction around the cell or on the surface of the cell, about which antigen prepared to be microscopically visible particles was distributed.
The techniques employed were the test tube method to study the on-cell-surface reaction and the slide glass method to observe the around-cell reaction. The latter reaction was thought to be more desirable than the former in respect to the specificity.
This method was tested in 126 cases of autoimmune hemolytic anemia (AIHA), systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), chronic thyroiditis and normal adults.
In the cases of AIHA, the erythorocyte of the patients themselves treated with bromelin or ficin were used as the antigen. For the cases of RA commercially avairable latex particles coated with denated human γ-globulin or tanned sheep erythorocytes coated with same antigen, for the cases of SLE latex particles coated with nucleoprotein, tanned sheep erythorocytes coated with DNA, or tanned llama erythorocytes coated with histone, and for the cases of chronic thyroiditis latex particles coated with thyroglobulin or tanned sheep erythorocytes coated with the same antigen, were used as the antigen.
Antibody producing cells were prepared from peripheral, blood lymph node and bone marrow. In an AIHA case was used spleen cells after the operation. Antibody producing cells in TC-199 were incubated at 37°C for 30 minutes and then antigen particles were added in the cell suspension. The mixture was kept at room temperature for 30 minutes shaking gently. A drop of the cell layer was taken from the sediment onto the slide glass, enclosed with a cover glass, and then observed with a phase contrast microscope. When latex particles covered more than 1/8 of the cell boundary or more than 4 coated erythorocytes adhered the cell, these cells were counted as the positive cell in which antibody was produced.
It is necessary to differenciate antibody producing cells from antibody fixing cells in the on-cell-surface reaction. The methods were discussed and tested.
The appearance rate of the auto-antibody producing cell by latex method was, 10 to 32 per cent in RA, and O to 12 per cent in SLE. These rate are higher than those which observed by the erythorocyte method. The latters were 1 to 3 per cent in peripheral lymphocytes and 2 to 10 per cent in the lymph node cells. This difference may be due to the fact that latex method is concerned partly with non-specific reaction. There was no direct orrelation between the serum antibody titer and the number of positive cells.
It has been shown that the positive reaction of SLE is specifically inhibited by anti-IgG serum (γ-chain specific) and that of RA by anti-IgM serum (μ-chain specific) These data suggest that antibody producing cells of SLE and RA are producing IgG and IgM antibody respectively.
This method is based on the virgin antibody, which was named and discussed by authorsin 1969.
It may be said that method has enabled us to identify the auto-antibody producing cell, though there are some problems yet to be resolved concerning specificity of the reaction.