Ensako, Katuhide

Fujiwara, Tadaaki

Nishiya, Koji

Ofuji, Tadashi

Yamana, Seizo

Yano, Keisuke

Okayama University Digital Information Repository

Studies on the population of human peripheral lymphocytes forming rosette with dog erythrocytes

Okayama University Scientific Achievement Repository

Acta Medica OkayamaVolume 28, Issue 5 1974 Article 7OCTOBER 1974Studies on the population of human peripherallymphocytes forming rosette with dogerythrocytesKoji Nishiya∗ Seizo Yamana† Keisuke Yano‡Katuhide Ensako∗∗ Tadaaki Fujiwara†† Tadashi Ofuji‡‡∗Okayama University,†Okayama University,‡Okayama University,∗∗Okayama University,††Okayama University,‡‡Okayama University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Acta Merl. Okayama 28, 373-375 (1974)-- BRIEF NOTE--STUDIES ON THE POPULATION OF HUMAN PERIPHERALLYMPHOCYTES·FORMING ROSETTE WITHDOG ERYTHROCYTESKoji NISHIYA, Seizo YAMANA, Keisuke YANO, Katuhide ENSAKo,Tadaaki FUJIWARA and Tadashi OFUJIDepartment of Internal Medicine, Okayama University Medical School,Okayama, Japan. (Director: Prof. T. Ojuji)Receivedjor publication, June 25, 1974In 1969, Dr. BACH (1) demonstrated that the lymphocytes in non-immunizedmice formed rosette with sheep red blood cells (SRBC). It is well known thata large population of human peripheral lymphocytes that form spontaneousrosettes with SRBC belongs to the thymus derived lymphocytes (T cell). Theevidence for this depends on that almost all the human thymocytes formrosettes, and it is blocked by a specific anti-thymocyte serum (2, 3). Themechanisms and the causes of rosette formation, however, are still obscure.Recent reports have demonstrated that a low but significant percentage ofhuman lymphocytes can bind the red blood cells of pig (4).We examined the spontaneous rosette formation between peripherallym-phocytes and red blood cells in various animals such as dogs, guinea pigs,rats, mice and human beings.Heparinized peripheral blood was obtained by heart puncture or veno-puncture, and the lymphocytes were separated using Conray-Ficoll gradientsedimentation method, and the 5 X 106/ml lymphocytes were suspended inHanks' balanced salt solution (Hanks' BSS) including 10% (vIv) fetal calfserum (FCS). Various kinds of red blood cells were also suspended in theidentical solution to make the final concentration to O. 5% (vIv) erythrocytesuspension. Equal volumes (0. 5 ml) of lymphocyte and red blood cell suspen-sion were mixed in a test tube (10 X 150 mm), centrifuged at 180 g. avo for5 min and incubated for 30 min at room temperature. After incubation, thesuspension was gently mixed at 33 r. p. m. for 5 min. One drop of the mixedsuspension was put on a slide glass, mounted by a cover slip, and erythrocyterosette-forming lymphocytes were counted under the light microscope. Lym-phocytes possessing three or more adherent erythrocytes were evaluated aspositive cells.As a result it was found that a low percentage rosette formation was be-tween guinea pig lymphocytes and rat erythrocytes (3.0%), dog lymphocytes3731Nishiya et al.: Studies on the population of human peripheral lymphocytes formingProduced by The Berkeley Electronic Press, 1974374 K.NISHIYA, S. YAMANA, K. YANO, K.ENSAKO, T.FuJIWARA and T.OFUJIand guinea pig erythrocytes (2.4%), human lymphocytes and mouse erythro-cytes (1. 7%). However, an increased number of rosette formation betweenhuman lymphocytes and dog erythrocytes (DRBC, 35. 1%) was observed.The intensity of this reaction seemed to be temperature-dependent, giving anoptimal binding at room temperature e. g. appropriately at 15°C.In addition, the population of DRBC rosette forming lymphocytes (DRBC-RFL) was compared with that of SRBC rosette forming lymphocytes (SRBGRFL). Namely, peripheral lymphocytes challenged with SRBC in advancewere divided equally into two test tubes. To one test tube, an equal volumeof SRBC suspension was added again and to the other, the DRBC suspension.The subsequent procedures were as previously described. The former (SRBC)experiment revealed the mean±SD=53. 6±9. 0% and the latter (DRBC),68. 3±9. 6% on the average of five preparations (P<O. 05). Similar experi-ments using DRBC rosette forming lymphocytes were performed, whichyielded nearly the same results as with SRBC rosette forming lymphocytes.We observed in the present experiments that both sheep and dog erythrocyteswould attach to the same lymphocyte. Therefore, we conclude that one groupof DRBC-RFL belongs to the same population as that of SRBC·RFL and othergroup is of a different population. As already mentioned, SRBC-RFL belongsto T cell, so that a major part of DRBC-RFL would be.long to the T cellpopulation, but it may be said that at least one group of the DRBC·RFL popu-lation might be different from SRBG-RFL population.TABLE I SPONTANEOUS" SRBC AND DRBC" ROSETTE FORMATION OFHUMAN PERIPHERAL LYMPHOCYTESHuman SRBC SRBC* SRBC** DRBC DRBC DRBClymphocyte + + + +SRBC DRBC DRBC SRBC------I 49.0 (%) 51.5 77.6 2.9 32.8 50.02 38.0 37.0 77.7 8.9 25.5 58.03 48.8 57.5 66.5 7.2 34.3 51. 74 51.5 62.4 68.2 12.8 35.0 58. 75 44.6 59.5 51.6 9.5 34.2 48.4Mean 46.4 53.6 68.3 8.3 32.4 53.4± ± ± ± ± ± ±SD 4. 7 9.0 9.6 3.2 3.5 4.2(P<0.05)* incubated first with SRBC, then repeated, indicating an increase of the rosette formingcells** incubated first with SRBC, then repeated using DRBC instead of SRBC, indicating anincrease of the rosette forming cells as well2Acta Medica Okayama, Vol. 28 [1974], Iss. 5, Art. 7http://escholarship.lib.okayama-u.ac.jp/amo/vol28/iss5/7Dog Red Cell-rosette Forming LymphocyteFig. I Rosette forming lymphocytesI. Human peripheral lymphocyte forming rosette with sheep red bloodcells. (X 1600)2. Human peripheral lymphocyte forming rosette with both sheep (smallone) and dog (large one) red blood cells. (x 1600)3. Human peripheral lymphocyte forming rosette only with dog redblood cells. (x 1600)REFERENCES3751. BACH, J. F., MULLER, J. Y. and DARDENNE, M.: In vivo specific antigen recognition byrosette forming cells. Nature 227, 1251, 19702. BACH, J. F., DORMONT, j., DARDENNE, M. and BALNER, H.: In vitro rosette inhibition byantihuman antilymphocyte serum. Transplantation 8, 265, 19693. BRAIN, P., GORDON, J. and WILLETTS, W. A.: Rosette formation by peripheral lympho-cytes. Clin. expo Immunol. 6, 681, 19704. LAY, W.H., MENDES, N.F., BIANCO, C. and NUSSENZWEIG, Y.: Binding of sheep red bloodcells to a large population of human lymphocytes. Nature 230, 531, 19693Nishiya et al.: Studies on the population of human peripheral lymphocytes formingProduced by The Berkeley Electronic Press, 1974

Studies on the population of human peripheral lymphocytes forming rosette with dog erythrocytes

Abstract

Similar works

Full text

Available Versions

Okayama University Digital Information Repository

Okayama University Scientific Achievement Repository