It has been demonstrated that by the mixed cultures in the presence of PHA the combination of those cells whose H-2 antigens differ from each other shows a higher rate and more significant difference of blastformation
than in the combination where non-H-2 antigens differ (Table 1). The blastformation observable in the combinations where non-H-2 histocompatibility antigens or sex.linked antigens are weaker, is not, so
marked as the difference seen of the blastformation in the case with H-2 isoantigens.
This in vitro lymphocyte stimulation test can be applied to the histocompatibility test in the combinations of strong H-2 antigens.</p

Kaneda, Shoken

Okayama University Scientific Achievement Repository

Acta Medica OkayamaVolume 23, Issue 3 1969 Article 3JUNE 1969Identification of mouse H-2 antigens by mixedlymphocyte culture in the presence of PHA. 3.Blastformation of mouse lymphocytesaccording to the difference in non- H-2antigensShoken Kaneda∗∗Okayama University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Identification of mouse H-2 antigens by mixedlymphocyte culture in the presence of PHA. 3.Blastformation of mouse lymphocytesaccording to the difference in non- H-2antigens∗Shoken KanedaAbstractIt has been demonstrated that by the mixed cultures in the presence of PHA the combinationof those cells whose H-2 antigens differ from each other shows a higher rate and more significantdifference of blastformation than in the combination where non-H-2 antigens differ (Table 1).The blastformation observable in the combinations where non-H-2 histocompatibility antigens orsex.linked antigens are weaker, is not, so marked as the difference seen of the blastformation inthe case with H-2 isoantigens. This in vitro lymphocyte stimulation test can be applied to thehistocompatibility test in the combinations of strong H-2 antigens.∗PMID: 4242842 [PubMed - indexed for MEDLINE] Copyright c©OKAYAMA UNIVERSITYMEDICAL SCHOOLActa Merl. Okayama 23, 203-207 (1969)IDENTIFICATION OF MOUSE H.2 ANTIGENS BY MIXEDLYMPHOCYTE CULTURE IN THE PRESENCE OF PHAIll. BLASTFORMATION OF MOUSE LYMHOCYTESACCORDING TO THE DIFFERENCE INNON. H-2 ANTIGENSShoken KANEDADepartment of Surgery, Okayama University Medical School, Okayama,Japan (Directc-r: Prof. S. Tanaka)Received for publication, March 20, 1969In the mixed cultures of mouse lymphocytes with different H-2 anti-gens conducted in such a way that only one cell group in the combinationof the two is made to react, it has been shown that the blastformation ofthese small lymphocytes results from lymphocytes reacting more potentlyupon the other lymphocytes having H-2 antigen lacking inthemselves, andthe lymphocyte stimulation test in vitro is an excellent one for the histo-compatibility test in making H-2 antigens of the two cell groups matchwith each other in order to identify the cells with a stronger H-2 antigens.However, supposing if the blastformation should occur in the same degreeas in the case against such antigens as non-H-2 histocompatibility antigenand sex-linked antigen which are the so·called weak histocompatibilityantigens, the blastformation in vitro can no longer be suitable test of hist-ocompatibility. In other words, such strains of lymphocytes that showstrong blastformation can no longer be excluded as an unsuitable recipient.By the mixed cultures of human peripheral blood lymphocytes ofclose blood relatives the blastformation is already shown to be weaker thanin the mixed cultures of similar lymphocytes of non-related persons (1).While there are dissenting opinions (2, 3) and no concrete genetic basis forit has been established yet, such a phenomenon seems to reflect one ofgenetic factors. We performed repeated mixed cultures of the mouselymphocytes with the so-called weak histocompatibility antigens, using theinbred mice whose gene is distinct but differing in their non-H-2 antigensor sex-linked antigen, though H-2 alleles are the same. As a result wefound clearly that the extent of blastformation is less marked in the casesof the lymphocytes with weaker histocompatibility antigens. This reportdeals with our findings on this study.2031Kaneda: Identification of mouse H-2 antigens by mixed lymphocyte cultureProduced by The Berkeley Electronic Press, 1969204 S. KANEDAMATERIALS AND METHODSThe methods and materials used are identical with those in the previouspaper, and they are omitted here.Lymphocytes: Axillary and cervical lymph nodes were obtained from A (H_2a )mice, C3H, CBA (H_2k ) mice all being adult mice of both sexes, and these lymphnodes were minced, strained through the filter, and lymphocytes so preparedserved as the materials.Medium: The culture medium was composed of TC-199+ YLE+ Hanks solu-tion -+- bovine serum, 5: 2 : 1 : 2 (vIv).Antibiotics used was penicillin (Takeda Pharmaceutical Co.).Culture methods: Both in the single and mixed cultures 100 x 104 cellslml wereused in each culture. The culture was conducted by the stationary method at37°C for 72 hours. Specimens were prepared at certain intervals, stained withMays-Giemsa stain, and the lymphocytes were classified according to their size:large 110 p.2 >, intermediate 110 p.2> > 56 p.2, and small 56 p.2,RESULTSIn every case the culture was carried out in the presence of 1 % PHA(vIv). The rate of blastformation (the appearance of large and inter.mediate cells) is as shown in Table 1. It was found that the rate of blast.formation was:Percentage of large and intermediate cells in mixed and unmixed culturesStrain combinationPercentage wIth phytohemagglutinin-MExperiment 2 3 4 5 AverageNo.A + C3H 27.4 40.2 3M.4 34.8 32.4 34.8CBA + C3H 17.2 30.6 22.2 19.6 20.2 22.0A'O + C3H'O 50.2 24.2 26.2 20.4 32.4 30.7A'O + C3H!f 26.6 30.0 21.0 34.2 28.3 28.0C3H o + C3H!f 18.5 16.4 14.4 18.8 20.4 17.7C3H'O 12.0 14.2 11.8 14.6 10.4 11.6A!f + C3H'O 39.4 32.0 29.4 37.6 33.3 34.3A!f + C3H!f 30.6 35.4 46.2 24.8 40.2 35.4A'O + A !f 20.2 24.4 25.6 17.6 20.8 21.7A!f 16.0 18.8 20.6 15.0 13.4 16.81) I n single strain culture 14. 2 % in average, 11. 6 % in the case ofC3H cells and 16.8 % with A cells. 2) Mixed culture in the combinationof the same strain but different sex: average was 19.7 %; 3) Hetero-2Acta Medica Okayama, Vol. 23 [1969], Iss. 3, Art. 3http://escholarship.lib.okayama-u.ac.jp/amo/vol23/iss3/3Identification of Mouse H-2 Antigens 205genous strain in the combination of the same sex: aveage was 33 %.4) Heterogenous strain in the combination of the different sex: averagewas 31.17 %.In other words, the blastformation was 1) < 2), showing it to be some·what greater in the combination of the same strain of opposite sex. Incomparing the results of 2), 3) and 4), the order of blastformation is 2) <3), 4), indicating that it is higher in the heterogenous combination. Therecan be recognized no difference in the blastformation between hetero-genous-same sex combination and heterogenous-opposite sex combination.Furthermore, in comparing the heterogenous mixed cultures themselves,the combination like (CBA + C3H) where the difference lies in somethingother than H·2 antigen, and in the combination of (A+C3H) where H-2differs, the rate of blastformation is higher in the combination where H-2antigens differ.SUMMARY AND DISCUSSIONIt has been demonstrated that blastformation is difficult to observe inthe combination where both groups of the cells in the mixed culture areof the so-called weak histocompatibility antigens and also the extent ofblastformation is slight in the case two groups have a marked differencein H.2 antigens such as A mouse lymphocytes with abundant H·2 isoanti.gens against C3H mouse lymphocytes with little H.2 isoantigens. Thesefacts seem to show gene-dosage effect. It is already demonstrated that theskingraft transplanted between mice whose antigen& differ at more thantwo weak H loci is rejected earlier than the skin transplanted between themice whose antigens differ only at one H locus. Even in the cases of micewith H.2 isoantigens such a dosage effect (additive effect) is observable(6, 7), GRAFF et ai. (5) performed skin graft transplantation experimentsby using various oncogenic resistant lines of mice (males and females) as wellas various combinations where antigens differed at 2, 3, 4, or weaker Hloci using non-H-2 or sex-linked histocompatibility antigens, and also thecombinations where antigens differed both at H-2 locus and non.H.2 locus,and observed the survival time of homo-skin grafts. They found that in.the combination where H.2 locus was the same but differed at non-H-2locus the weak H locus showed additive effect, and the greater thedifference the greater was the transplantation immunity, and as such adifference grew more marked, the skingraft is rejected around 12 days,just as in the case of skingraft with antigens differing at H locus.The potency of histocompatibility antigens depends upon the extent3Kaneda: Identification of mouse H-2 antigens by mixed lymphocyte cultureProduced by The Berkeley Electronic Press, 1969206 S. KANEOAof immune response induced by the antigen, and in the skin homograftsbetween two strains whose antigens differ at H.2 locus the survival of theskin grafts is 8--10 days, but in the cases where antigens differ only at H.land H.3 (H.2 being the same), such skin graft survives several tens oreven 100 days. KLEIN (8) has demonstrated that the majority of individualnon.H.2 antigens act as a weak antigen as judged from the numbers ofsurvival days of the skin homografts. It may be said that a strong histo.compatiblity locus means a complex locus that controls many isoantigens,and the weak locus is the site where a small number of isoantigens aregathered.It appears that for the induction of blastformation in lymphocytesthere should be a difference in a considerable number of histocompatibilityantigens as well as the difference in more than one H.2 isoantigens. Aspostulated by the so·called clonal theory of BURNET, the presence of lymph.ocytes which are destined to give rise to blastformation in response toantigens, brings about additive effect, and this phenomenon may beinterpreted as the stronger the histocompatibility antigens of the lympho.cytes the greater is the rate of blastformation, and conversely, the weakeris such histocompatibility antigens, the lesser the blastformation. The factthat, contrary to expectation, the combination of non.H.2 antigen yieldedthe strongest response in the mixed cultures of mouse lymphocytes asobserved by FES1'ENSTEIN (9) using the C J4 .thymidine.incorporation as thecriterion, may somehow be explained by the gene.dosage effect.In their mixed cultures of human lymphocytes in which there was aderangement of 4 antigens, 1, 2, 3, and 7 belonging to the major histo.compatibility loci and those without such a derangement, I VANJI et ai. (l0)observed a higher rate of blastformation in those with the derangementbut in those without such derangement, that is, the blastformation corres.ponding to those of strong antigens and the combination of differentlymphocytes with weak antigens, the blastformation was less. Amongnumbers of histocompatbility testings the most feasible ones that do notsensitize the recipient nor give pain to either recipient or donor are twotests: namely, the mixed lymphocyte culture test and the serological testby the use of human antiserum. However, since the results by these twotests concert with one another (10, 11), the mixed lymphocyte culture test,provided that the major histocompatibility antigens as well as the weakhistocompatibility antigens are contained numerously, would serve as thecriterion for determining the blastforrnation, hence it would be sufficientlyuseful as an exclusion test of the donor.4Acta Medica Okayama, Vol. 23 [1969], Iss. 3, Art. 3http://escholarship.lib.okayama-u.ac.jp/amo/vol23/iss3/3Identification of Mouse H-2 AntigensCONCLUSION207I t has been demonstrated that by the mixed cultures in the presenceof PHA the combination of those cells whose H.2 antigens differ fromeach other shows a higher rate and more significant difference of blastfor-mation than in the combination where non-H.2 antigens differ (Table 1).The blastformation observable in the combinations where non.H.2histocompatibility antigens or sex.linked antigens are weaker, is not, somarked as the difference seen of the blastformation in the case with H-2isoantigens.This in vitro lymphocyte stimulation test can be applied to the histo.compatibility test in the combinations of strong H.2 antigens.REFERENCES1. CHALMERS, D. G. et al.: Immunologically stimulated human peripheral blood lympho-cytes in vitro. Int. Arch. Allergy 30, 177, 19662. RUBIN, A. C. et al.: Histocompatibility and immunologic competence in renal homo-transplantation. Science 143, 815, 19643. ELVES, M. W. and ISRAELS, M. G. G.: Lymphocyte transformation in cultures ofmixed leucocytes. A possile test of histocompatibility. Lancet 1, 1184, 19654. MCKHANN, C. F.: Additive effect of multiple weak incompatibilities in transplantationimmunity. Nature 201, 937, 19645. GRAFF, R. J., SILVERS, W. K., BILLINGHAM, A. E., HILDEMANN, W. H. and SNELL, G.0.: The effect of histocompatibility antigens. Transplantation 4, 605, 19666. KOKUMAI, Y.: Humoral and cellular reactions against the transplanted cells. Acta Me-dicinae Okayama 20, 235, 19667. HILDEMANN, W. H. and MEDAWAR, P. B.: Relationship between the transplantationimmunity and formation of humoral isoantibodies in mice. Immunology 2, 44, 19598. KLEIN, J.: Strength of Histpcompatibility Genes in Mice in Histocompatibility Testing.(CURTORI, E. S., MATTIUZ, P. L. and TOST, P. M,) Munksgard, Copenhagen pp 21-299. FERSENSTEIN, H.: Antigenic strength investigated by mixed culture of allogenic mousespleen cells. Ann. N. Y. Acad. Sci. 129, 567, 196610. IVANYI, D., RYCHLIKOVA, M., SASPOTES, M., IVANYI, P., DAUSSET, J.: Leukocyte anti-gens and the mixed lymphocyte culture reaction. Vox. Sanguinis 12, 186, 1967II. BACH, F H. and AMOS, D. B.: Hu-I: Major histocompatibility locus in man. Science156, 1506, 19675Kaneda: Identification of mouse H-2 antigens by mixed lymphocyte cultureProduced by The Berkeley Electronic Press, 1969

Identification of mouse H-2 antigens by mixed lymphocyte culture in the presence of PHA. 3. Blastformation of mouse lymphocytes according to the difference in non- H-2 antigens

In the mixed tissue culture of mouse lymphocytes with addition of PHA the rate of the appearance of large and intermediate cells increases markedly, but which side of the two cell groups have reacted stronger remains obscure. In order to solve this problem, mixed cultures were
conducted in such a way that only one cell group of the two would react. Namely, one cell group was exposed to C0 6).irradiation (Table 2) prior to the culture and cultured with another viable cell group (FJ test group, Table 2) to see the percentage of the appearance of large and intermediate cells. Simultaneously, the skin homograft from respective donor mouse was transplanted to each other and the survival days of each skin graft were compared. As a result it has been shown that the percentage of
blastformation and the survival time of the skin transplant in each group prove to be in an inverse relation. The results of these mixed cultures indicate that the extent of blast. formation reflects significantly the difference in B-2 histocompatibility antigens.</p

Acta Medica OkayamaVolume 23, Issue 2 1969 Article 2APRIL 1969Identification of mouse H-2 antigens by mixedlymphocyte culture in the presence of PHA. II.Blastformation of mouse lymphocytes inculture according to the difference in H-2antigensShoken Kaneda∗∗Okayama University,Copyright c©1999 OKAYAMA UNIVERSITY MEDICAL SCHOOL. All rights reserved.Identification of mouse H-2 antigens by mixedlymphocyte culture in the presence of PHA. II.Blastformation of mouse lymphocytes inculture according to the difference in H-2antigens∗Shoken KanedaAbstractIn the mixed tissue culture of mouse lymphocytes with addition of PHA the rate of the ap-pearance of large and intermediate cells increases markedly, but which side of the two cell groupshave reacted stronger remains obscure. In order to solve this problem, mixed cultures were con-ducted in such a way that only one cell group of the two would react. Namely, one cell group wasexposed to C0 6).irradiation (Table 2) prior to the culture and cultured with another viable cellgroup (FJ test group, Table 2) to see the percentage of the appearance of large and intermediatecells. Simultaneously, the skin homograft from respective donor mouse was transplanted to eachother and the survival days of each skin graft were compared. As a result it has been shown thatthe percentage of blastformation and the survival time of the skin transplant in each group proveto be in an inverse relation. The results of these mixed cultures indicate that the extent of blast.formation reflects significantly the difference in B-2 histocompatibility antigens.∗PMID: 4242309 [PubMed - indexed for MEDLINE] PMID: 4242309 [PubMed - indexed forMEDLINE] Copyright c©OKAYAMA UNIVERSITY MEDICAL SCHOOLActa Merl. Okayama 23, 89-94 (1969)IDENTIFICATION OF MOUSE H.2 ANTIGENS BY MIXEDLYMPHOCYTE CULTURE IN THE PRESENCE OF PHAII. BLASTFORMATION OF MOUSE LYMPHOCYTESIN CULTURE ACCORDING TO THEDIFFERENCE IN H.2 ANTIGENSShoken KANEDADepartment of Surgery, Okayama University Medical School, Okayama,Japan (Director: Prof. S. Tanaka)Received for publication, January 12, 1969It has been demonstrated that, when mouse lymphocytes are culturedIII the medium containing PHA, either in single or mixed culture withother cells, the rate of large and intermediate cell appearance is markedlyenhanced. However, when these lymphocytes are cultured in variouscombinations with other cells in the absence of PHA, there can be obser·ved hardly any significant difference in the rate of blastformation betweenvarious combinations, while there is a distinct difference in the presenceof PHA. This fact suggests that the H-2 antigen histocompatibility differ.ence is made more marked on addition of PHA. It remains, however,still obscure which side of the cell groups in the combination respondsmore markedly. In order to solve this question, a series of mixed cellcultures were conducted under such conditions that only one group of thecells in the two·group combination was made to react. Namely, onegroup of the lymphocytes (A or C3H cells) was previously irradiated withCOOD to inhibit mitosis, and then one irradiated group was cultured withthe other non.irradiated group: either in the combination of irradiatedA + C3H cells or irradiated C3H + non.irradiated A cells. Simultaneously,F1 mouse lymphocytes were cultured with their parent lymphocytes (as theF1 test); F1 + A or F1 + C3H. These mixed cultures were conducted inorder to see what relation would blastformation show with the survivaltime of the skin homograft transplanted.MATERIALS AND METHODSThe methods employed here are identical with those used in Part 1.Lymphocyte: Lymphocytes are isolated from cervical and axillary lymphnodes of A mice (H_2a), C3H mice (H_2k ), and (A X C3H) F 1 mice, both frommale and female adult mice, as in the previous experiment.891Kaneda: Identification of mouse H-2 antigens by mixed lymphocyte cultureProduced by The Berkeley Electronic Press, 196990 S. KANEDAIrradiation of the cells with C0 60: The irradiation is done under the condi.tions of 10'R of C06); 200 kv; 25 mA; 0.54/--,+0.5 Al filter; focal distance 20cm; time 22' 22". After confirming the death of the lymphocytes by supra.vital staining (l, 2), the mixed cell cultures are carried out.Culture medium: The medium is composed of TC-199 + YLE +- Hanks solu.tion + bovine serum, 5: 2 : 1 : 2 (vIv).PHA: One vial of hemagglutinin M (Difco Laboratories, Detroit, Michi.gan, U. S. A.) is dissolved in 5 ml of the medium. Antibiotics used is penicillin(Takeda Pharmaceutical K. K.).Skin tramplantation: The skingraft in the direction of F1- A, and Fl-C3Hin full thickness is transplanted on the back of the mice. The rejection of theskingraft is determined macroscopically, and each of these two groups iscomposed of 20 animals.Culture methods: In both single and mixed cultures 100 x 104 lymphocytesare used each time. The static cultures are carried out at 37°C for 72 hours,and the specimens are taken at certain intervals, stained with May-Giemsastain, and the cells are classified as large, intermediate and small lymphocytes(3, 4).RESULTS1. The mixed culture of Coon-irradiated lymphocytes and non-irradiated normallymphocytes :In the combination of irradiated A cells + normal nonirradiated C3Hcells without addition of PHA the rate of appearance of large and inter-mediate cells was 7.0 %, while in the reverse combination of non-irradiatedA cells + irradiated C3H cells, the rate proved to be 5.35 %, showinghardly any significant difference between them. In the presence of PHA,however, the rate of appearance in the former combination was 27.95 %and 18.1 % in the latter, indicating a distinct difference (p= 0.05) betweenthe two groups.2. llfixed cultures of F'r cell +parent cells:In the combination of A+ F1 cells without PHA the rate of large-intermediate appearance (blastformation) was 7.45 % while it was 9.70 %in the combination of C3H + Fp revealing hardly any significant differencebetween the two combinations. However, on the addition of PHA, therate of blastformation in the former combination was 19.3 % and 31.85 %in the latter, showing a significant difference (p=0.05).3. Survival time of the homograft of the skin from F; mice to parent mice:In the transplantation of the skin graft from F j mice to the parentmice, the survival days proved to be 14+ 1. 22 days in average, and in theF j to C3H mice transplantation it was 9.7 + 0.28 days in average, reveal-2Acta Medica Okayama, Vol. 23 [1969], Iss. 2, Art. 2http://escholarship.lib.okayama-u.ac.jp/amo/vol23/iss2/2Identification of Mouse H-2 Antigens in Mixed Caltures with PHA 91ing a significant difference (p=O.Ol) between the two sets (Table 1).Table 1. Percentage of Large and Intermediate Cells in Mixed Culturesof F, Cells and Parent CellsExp. Nos. Strain Combination Without PHA With PHAA + F, 8.2 % 15.4 %C3H + F) 7.6 % 32.2 %2 A +F) 7.2 % 20.6 %C3H + F) 12.8 % 34.0 %3A + Fl 10.4 % 22.8 %C:jH + F, 12.0 % 40.0 %A + F, 4.0 % 18.4 %4C3H + F) 6.4 % 21.2 %A + F, 7.45 % 19.30 %Average: C3H + F, 9.70 % 31.85 %p = 0.3 P = 0.05SUMMARY AND DISCUSSIONIt is said that 97 per cent of mouse lymphocytes are small lymphocytes(4) and hence those cells that show blastformation in tissue culture arethought to be small lymphocytes (5-11).It is also considered that the greater is the genetic difference betweenthe lymphocytes in mixed culture, the greater is the extent of blastform-ation. However, the lymphocytes irradiated with a large dose of C0 63 andF) cells would theoretically not respond to the target cells, lymphocytesand their parent cells (12) On the other hand, the findings that theblastformation in the combination of irradiated A cells + normal C3Hcells is greater than in the reverve combination of normal A cells + irradi-ated C3H cells, and that in F) cells + C3H cells is greater than in thecombination of F) cells + A cells indicate that in either case, C3H cellswith less histocompatibility antigens react more potently on H-2" antigensof A cells possessing more abundant H-2 antigenic components. Further-more, since the in vivo result shows that the homograft of skin transplantedfrom F1 to C3H mice is rejected earlier than that transplanted from F] to Amice and this phenomenon is in a reverse correlation with the rate ofblastformation in the mixed culture of lymphocytes, it suggests that the invitro mixed lymphocyte culture test can be used for the histocompatibility3Kaneda: Identification of mouse H-2 antigens by mixed lymphocyte cultureProduced by The Berkeley Electronic Press, 196992 S. KANEDAtesting. It follows then, if we select from various prospective donor lym.phocytes and treat them by irradiation or by some other means wherebythey are made unable to undergo blastformation and by selecting thosewith the least blastformation we can have a proper donor. Hence it isobvious that the skin graft from (A X C3H) F1 donor is better accepted byA mice than by C3H mice.Since non.H·2 antigens apart from the H-2 histocompatibility shoulddiffer between A mice and C3H mice, it seems rather unreasonable tolimit the blastformation and the survival of homotransplantation of theskin only to the difference in the H-2 antigens. Therefore, it is necessaryto study to what extent the blastformation differs owing to the differencein non·H-2 antigens.There are reports dealing with the mutual relationship between thedifference in mouse H-2 antigens and blastformation as by DUTTON andFESTENSTEIN, all these studies are done on the basis of the incorporationof labeled thymidine into the nucleus at the time of mitosis. DUTTON (13)in his mixed cultures of spleen cells from various mice of in bred strainsconducted for 54 hours reported that quite variations were found in theincorporation of labeled thymidine. FESTENSTEIN (14) likewise conductedmixed cultures of spleen cells from various inbred mice for 5 days to deter-mine blastformation of the culture cells on the basis of the extent of CJ4•thymidine incorporation, but reported that there could be observed nosignificant correlation between the difference in H.2 antigens and theextent of response. However, none of them used PHA in their culturemedium, hence the resultant blastformation was poor as to reflect suffici.ently the differences in the H-2 antigens. It seems obvious that PHAenhances the specific reaction on the histocompatibility antigen of thecells.CONCLUSIONIn the mixed tissue culture of mouse lymphocytes with addition ofPHA the rate of the appearance of large and intermediate cells increasesmarkedly, but which side of the two cell groups have reacted strongerremains obscure. In order to solve this problem, mixed cultures wereconducted in such a way that only one cell group of the two would react.Namely, one cell group was exposed to C0 6).irradiation (Table 2) prior tothe culture and cultured with another viable cell group (FJ test group,Table 2) to see the percentage of the appearance of large and intermediatecells. Simultaneously, the skin homograft from respective donor mouse4Acta Medica Okayama, Vol. 23 [1969], Iss. 2, Art. 2http://escholarship.lib.okayama-u.ac.jp/amo/vol23/iss2/2Identification of Mouse H-2 Antiens in Mixed Caltures with PHA 93was transplanted to each other and the survival days of each skin graftwere compared. As a result it has been shown that the percentage ofblastformation and the survival time of the skin transplant in each groupprove to be in an inverse relation.The results of these mixed cultures indicate that the extent of blast.formation reflects significantly the difference in B-2 histocompatibilityantigens.Table 2. Percentage of Large and Intermediate Cells in Mixed Culturesof Irradiated Cells and Untreated Allogeneic CellsExp. Nos.234AverageStrain Combination Without PHA With PHAjAI+ CaH 7.0 % 26.2 %A + !eaHl 4.2 % 20.2 %IAI+ CaH 6.2 % 21.0 %A + ICaH! 8.0 % 15.4 %IAI+ CaH 8.2 % 20.4 %A + ICaH! 6.2 % 12.2 %IAI+ CaH 6.6 % 34.2 %A + ICaHI 3.0 % 24.6 %W+ CaH 7.00 % 27.95 %A + ICaHI 5.35 % 18.10 %p=0.2 p=0.051=1: denotes irradiated cells.REFERENCES1. MCLIMANs, W. F., DAVID, E. V., GLOVER, F. L. and PAKE, G. W.: j. lmmunol. 79,428, 19572. SCHREK, R.: A method for counting the viable cells in normal and malignant cell sus-pensions. Amer. j. Cancer 28, 389, 19363. KUROYANAGI, T.: Thymus and immunity, Nihon Rinsho (japan Clinical Medicine) 22,2359, 19644. HANAOKA, M.: Lymphatic thymus systems and immunity. Nikketsu-Kaishi 27, 155, 19645. BURNET, M.: Role of the thymus and related organs in immunity. BJith. Med. j. II5380, 807, 19626. MILLER, I. F. A. D. et al.: Immuuo1ogica1 significance of the thymus in "Advances inImmunology" Academic Press. New York & London. 2, p. 111, 19627. GOOD, R. A. et al.: Outogeny and phylogeny of adaptive immunity in "Advances inImmunology" Academic Press. New York & London. 4, p.l, 19648. MILLER, ]. F. A. P.: Immunity in foetus and newborn. Brith. Med. Bull. 22, I, 21,5Kaneda: Identification of mouse H-2 antigens by mixed lymphocyte cultureProduced by The Berkeley Electronic Press, 196994 S. KANEDA19669. Special Issue: Various basic problems of thymus. Saishin Igaku 20, 2634, 196510. Special Issue. Lymph nodes and basic problems. Saishin Igaku 21, 1143, 196611. HANAOKA, M.: Antigen-formin cells. Gendai-no-SeibutJugaku (Modern Physiology) 5, 161,196612. HIRSCHHORN, K.: Lymphocyte in vitro alteration and response in immunopathology, VIInternational Symposium 1965, ed. P. GRABEN and D. A. MIESCHER, SCHWABE, Co.p. 157, 196613. DUTTON, R. W.: j. Exp. Med. 122, 759, 196514. FESTENSTEIN, H.: Ann. N. Y. Acad. Sc. 129, 567, 19666Acta Medica Okayama, Vol. 23 [1969], Iss. 2, Art. 2http://escholarship.lib.okayama-u.ac.jp/amo/vol23/iss2/2

Identification of mouse H-2 antigens by mixed lymphocyte culture in the presence of PHA. II. Blastformation of mouse lymphocytes in culture according to the difference in H-2 antigens

Identification of mouse H-2 antigens by mixed lymphocyte culture in the presence of PHA. 3. Blastformation of mouse lymphocytes according to the difference in non- H-2 antigens

Abstract

Similar works

Full text

Available Versions

Okayama University Scientific Achievement Repository

Okayama University Scientific Achievement Repository

Okayama University Digital Information Repository

Okayama University Digital Information Repository