A Japanese family, in which three members (propositus and his two sibs) are of phenotype P₂^k, and the frequency of P^k in Japanese population are described.
The family (Fig. 1). The propositus (II-6) is a 38-year-old healthy donor who comes from Iwate Prefecture in the northern part of Japan. His parents (I-1 and I-2) are first cousins. Two (II-5 and II-9) of five sibs of the propositus were also P₂^k and the other members of the family were P₂. Anti-P in the serum from the three P₂^k members was mainly IgG and partly IgM, though they had no history of blood transfusion.
The P₂^k members were tested for the 20 immunogenetic and biochemical systems (Table 1) and it was found that a gene responsible for P^k is independent of the loci for ABO, MNSs, Rh, Kidd, Hp, Gc, ACP, PGM₁ and GPT.
Frequency of P^k. The frepuency of the phenotype P^k was studied by two different ways.
1) During the last ten years, we have found three examples of P^k for the accompanying anti-P in testing over 270, 000 Japanese donors and pregnant women and their husbands, a frequency of 0.00001 (0.001%) with a upper limit (95% certainty) of 0.00002 (0.002%).
2) A total of 7 P^k propositi, including one reported in this paper, have so far been found in Japan (Hayashida, 1968; Yamaguchi et al., 1974; Furukawa et al., 1974; Furukawa and Kishi, 1976; Nakajima and Yokota, 1977). Four of the 7 propositi are the offspring of first cousin marriage: rate of the first cousin marriages is 57% and it raises to 71% if second cousins are included. According to Imaizumi et al. (1975), the mean rate of first cousin marriages in Japan until 1957 is about 5% (inbreeding coefficient=0.004).
By putting these values into the Dahlberg's formula, the frequency of gene responsible for P^k was estimated at 0.0026-0.0013 and that of phenotype P^k at 0.00017-0.000007 or 1 in 60 to 140 thousand. These figures are almost in agreement with those estimated in the population studies mentioned above. The rate of first cousin marriages in Japan has, however, shown a rapid decrease in recent two decades, showing about 1% or less (Imaizumi et al., 1975). Thus, the frequency of phenotype P^k in young people might be much lower than the above-estimated values.

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Here reported the results of the studies on the HLA antigenic difference between lymphocytes and granulocytes by using lymphocyte cytotoxicity test, lymphocyte agglutination test, granulocyte cytotoxicity test and granulocyte agglutination test. The cytotoxicity test was carried out by the NIH method and the agglutination test by van Rood's microagglutination method. The antisera used were HLA-A2, A9, BW40, multispecific and negative and positive control. Antisera were absorbed with the cells for 40min. at 37°C.
1) Cytotoxicity Test
Lymphocytes often reacted to the HLA antisera absorbed with granulocytes.
2) Agglutination Test
Antisera absorbed with each corresponding granulocytes, lymphocytes or total leukocytes almost completely failed agglutinate lymphocytes or granulocytes, except for a weak reaction in one case.
3) In order to compare the sensitiveness of agglutination test with that of cytotoxicity test, both tests were carried out using two-fold diluted control positive serum. The results revealed that the cytotoxicity was more remarkable in lymphocytes than in granulocytes.
The sensitivity of the leukocyte agglutination test was similar to that of lymphocyte cytotoxicity test, while granulocyte and lymphocyte agglutination tests showed lower sensitiveness.
4) Lymphocyte and granulocyte cytotoxicity tests as well as lymphocyte and granulocyte agglutination tests were carried out in members of 4 families using HLA sera of Terasaki series obtained from Behring Company. Lymphocyte cytotoxicity test and lymphocyte agglutination test showed similar patterns in each family, while granulocyte cytotoxicity test showed somewhat different patterns. Granulocyte agglutination test showed similar or different patterns compared with lymphocyte cytotoxicity test. When the parents were supposed to have the genes A, B, C and D, antigenic inheritance to descendants was recognized, but lymphocyte agglutination test, granulocyte cytotoxicity test and granulocyte agglutination test showed some discrepancy from lymphocyte cytotoxicity test. These results obtained revealed some antigenic difference between lymphocytes and granulocytes.

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Practical trials have been undertaken by the Antiglobulin Working Party of the ICSH/ISBT Joint Expert Panel for the standardization of antiglobulin reagent for use in compatibility test. The results obtained by the participants from our country are reviewed with special reference to the currently effective Japanese Minimum Requirements.
Internationally agreed specifications for antiglobulin sera will be of great help in reviewing and possibly revising our national Requirements. On the other hand, until an agreement could be attained by all the participants on the purpose of designating an international standard or reference preparation and on the function to be played thereby, the discussion on the selection of a candidate for the international standard will hardly find its final solution.

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