Fundamental studies on the peripheral leucocyte culture and its clinical application

With an improved method of tissue culture of peripheral leucocytes of our own design, the authors carried out systematic observations on the peripheral leucocyte culture from the normal, various kinds of leukemias, leukemoid reactions, and hypoplastic anemia. As for the culture method we have devised a method of silicon oil coating on the blood containers, a method which will least affect the cell function as compared with the conventional culture method. As the results we have found that the tissue growth in the case of peripheral leucocyte culture of normal persons ceases after six-hour culture and also we have recognized a peculiar finding, a growth like a corona-shape, in which an empty space appears in the inner part of growth area along with the lapse of time. In every leukemic case, without presenting growth area like a corona, the cell density is high and the outer zone of growth area becomes sharply demarcated as in the case with bone-marrow culture and also the growth continues even after 12 hours. At this instance we have noticed many mitoses of immature cells. Moreover, the maturation of immature cells has been observed and it has also been possible to distinguish to what type these immature cells belonged. In the leukemoid reaction no growth pattern Characteristic to the leukemia can be recognized, and at a glance it can easily be differentiated from the leukemia. In the case of hypoplastic anemia although the growth area is like that of the normal presenting a corona-like shape, the cell density is lower and the function of leucccytes is lesser than the normal. From these results we believe that the peripheral leucocyte culture is clinically useful.</p

マイクロサテライトDNAタケイジョウホウ ニ モトヅク リュウジンジドリ ノ イデンテキタヨウセイ

龍神地鶏は和歌山県の旧龍神村(現在の田辺市)で少数が維持されている集団であり,同地で古くから飼養されているものである。1994年には村内で30数羽が飼養されていたが,近年では個体数が減少し,遺伝的多様性の減少が懸念されている。そこで本研究では1994年および2007年に採血された龍神地鶏(1994年12羽,2007年2集団各18羽,7羽)について,ISAG/FAO推奨の30座位のマイクロサテライトマーカーを用いて遺伝的多様性の経時的な比較と他の日本鶏品種との遺伝的類縁関係を明らかにすることを目的とした。龍神地鶏3集団において30座位中12座位で多型が認められず,5座位で対立遺伝子の消失が認められた。さらに6座位においては遺伝子頻度0.5以上の主要な対立遺伝子が変化していた。その他の座位の対立遺伝子数は2から3の範囲であった。龍神地鶏各集団の平均対立遺伝子数およびヘテロ接合体率は既報の他の日本鶏品種よりも低い値を示した。次に,日本鶏品種内における龍神地鶏の遺伝的な位置を明確にするため,他品種の解析データを加えてD^A遺伝距離にもとづく近隣結合系統樹を作成した。その結果,龍神地鶏は比較に用いたどの品種ともクラスターを形成せず,高いブートストラップ値で他の品種から分かれる結果となった。以上の結果より,龍神地鶏は地域に固有の品種である一方,小集団で長く維持されてきたため近交がすすみ,遺伝的多様性が低くなった集団であると考えられた。今後この品種を維持するためには,現在残されている2つの集団のみならず,県の試験場等を含めて十分な集団サイズを確保し,集団間の系統的維持が必要であると考えられた。Ryujin-Jidori is one of the Japanese native chicken breeds kept in Ryujin village in Wakayama prefecture. Recently, a reduction in population size and in genetic diversity were detected in this breed. In this study, ISAG/FAO recommended 30 microsatellite markers be used to investigate temporal change in genetic diversity and phylogenetic relationships among Ryujin-Jidori and other Japanese native chicken breeds. The Ryujin-Jidori samples were collected in 1994 (n=12) and 2007 (two populations, n=18, 7), respectively. In this analysis, twelve monomorphic loci were observed. Allele loss at five loci and shift of major allele (allele frequency>0.5) at six loci were also observed. In other loci, number of alleles ranged from two to three. Mean number of alleles (MNA) and average expected heterozygosity (H^E) of Ryujin-Jidori populations were lower than previously reported in other Japanese native chicken breeds. No significant difference in MNA was observed between the two Ryujin-Jidori populations. However, a temporal decrease over time in H^E was observed. On the neighbor joining dendrogram based on D^A genetic distance, the Ryujin-Jidori populations were sepa- rated from all other breeds (Gifu-Jidori, Tosa-Jidori, Shokoku and Shamo) with robust bootstrap value. Therefore, it is suggested that Ryujin-Jidori is a unique and valuable genetic resource for Wakayama prefecture. However, to keep Ryujin-Jidori in small population could lead to an affect of genetic drift, intensify inbreeding and decrease genetic diversity