The invention relates to a treatment method of human peripheral blood T lymphocyte, animmunogen preparation, a preparation method of antiserum, and application. T lymphocytes are separated and purified by using a waste leukocyte filter disc, multiplication culture is carried out, and antigen surface marker identification comparison and activity comparison are carried out on the separated T lymphocytes and the cultured T lymphocytes; the high-activity T lymphocytes subjected to multiplication culture are used as an immunogen, antiserum is prepared by using a healthy pig, the titer after immunization is evaluated by using results of an E rose ring inhibition experiment and a cytotoxicity experiment, and formulating an immunization program, so as to prepare a large amount of antiserum; and anti-plasma is collected for plasma separation, and the plasma separation is used for preparation of ALG products. According to the invention, the limitation problem that fresh whole blood needs to be collected every time in the titer verification of the anti-human T lymphocyte immune globulin preparation raw material and the titer verification of the finished product can be solved, and meanwhile, the bottleneck problem of the anti-human T lymphocyte immune globulin raw material can also be solved.