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IN VITRO STUDIES OF PRESENT DOMINGO LUIs ON MUSCOLO, the Department of In vitro Orthopaedic studies cytotoxic Preliminary transplantation histocompatibility there are bone cells. branes attention of bone no reports have not has on cells been the antigenic been isolated components on from present in bone surface antigens seem to of bone cell great number the that generate play memof immune a key as bone Muscobo, agement albotranspbantation role (Ottolenghi in 1972; Kawai and Ray 1976), immunological of skeletal sarcomas (Invernizzi and 1975), and possible susceptibility diseases (McDevitt also in clarifying help disorders The and to some Bodmer the 1974), aetiobogy of those suspected of being related present study was undertaken presence of major antigens on bone in with vitro disparate mine if cells. individuals they carry lymphocytes. antiserum, plantation Isolated bone cells lymphocytes bone containing antigens, might the cells were antibodies in order to were (from cultured genetically species) to deterof stimulating exposed against major transstudy their antigenic MATERIALS AND METHODS Lewis, Brown-Norway were Bethesda, and each, the others purchased Shinya Robert from equivalent Domingo Luis and Buffalo adult Microbiological of human ‘1 . I). Niuseolo. Aires. male allogeneic lymphocyte hand, for the presence . HL-A I ntitute (Palm 1971). of Orthopaedics with and University of Illinois of transplantation lymphocytes On the other and exposed to their antigenic profile. antigens in an active bone cells of serologically were killed by defined (SD) absorbed sera suggest “sharing” of The relevance of the surface antigens susceptibility to various orthopaedic (Bronwill-Gilbings, Rochester, New York) fitted with ing marrow per cent cells, and transferred dipotassium to flasks salt of EDTA vated (56 serum in 20 milbilitres with 5 per cent degrees Celsius for of TC and thirty containing 1.5-1.7 cent heat macti- 5 per minutes) medium 199. autobogous The flasks were shaken in water gassed CO2 in air, sealed, and bath at 37 degrees Celsius for eighteen hOurs. After decalcification, the slices were washed, ded in flasks I, Type shaken digested. viability containing two The determined 60-70 per cent). haemocytometer 8x10xml). Fig. for resuspen- milligram of collagenase 150 units/mg) in 10 millilitres of medium, and hours or more until they were completely isolated bone cells were washed twice and by the trypan blue exclusion test (usually The cell concentration was determined by and phase contrast microscopy (normally The isolated 3) or were 2-3 cells bipolar showed multiple delicate processes with long unbranched processes as described. Cellular in vitro experiments When two populations of lymphoid cells from individuals genetically different at the major histocompatibility locus are cultured together in vitro, an interaction between the cells takes place and lymphocyte proliferation occurs. This reaction, called mixed lymphocyte culture (MLC), reflects antiments, differences mixed bating between lymphocyte-bone to determine lymphocyte the cells. In the present cell cultures (MLBCC) if bone cells proliferation are when experiwere capable cultured of stimutogether in vitro. With established Iraumatologv, that purpose (Fig. 1): Italian hospital. in mind the 1) lymphocytes U niver..itv of following cultures from Lewis rats+ Iienos Ares. Potoi were lym42 I 5. M.D.. Ray. ME). [)epartment of Orthopaedic Surgery. , Phi).. Departhient of Orthopaedic Yarnaguchi Surgery. University. University School of of Medicine. Illinois, 54() South lJhe \Vood (.atv. 75 Japan. Street. (‘hieago. IlIinoi #{241}l2. U.S.A. Requests 342 for a diamond impregnated blade. The slices were dipped into distilled sterile water for two seconds in order to byse any remain- established Associates, The ArgentIna. Kawal, D. Fisher Maryland. These strains of rats are highly inbred with the exception of Lewis and Fisher, differs from at the major histocompatibibity locus of the rat Ag-B-the Buenos (BN), Medicine, to determine stimulating genic Rats and JAPAN the presence antigens, previously to cytotoxic UBE, Isolation of bone cells The technique previously described by Bard, Dickens, Edwards and Smith (1974) was used with minor modifications. Soft tissues, periosteum and cartilage were removed from femora and tibiae and the marrow was removed by washing with TC medium 199 (Difco, Detroit, Michigan). Bone slices of approximately 150 micrometres were cut under continuous irrigation using a thin sectioning machine ( profile. rats with (Sigma orthopaedic (histocompatibility) within the same antigens capable Also they to auto-immunity. to investigate transplantation albogeneic manParmiani orthopaedic and of allotransplantation, upon KAWAI, to investigate cultured RAT OF AMERICA School transplantation may not express different biological phenomena (Bach, Bach and Sondel 1976), and it is likely that attempts to define those antigens on bone cells could be relevant to clinical situations such SHINYA undertaken albografts, THE STATES Lincoln were focused and/or structures antigens Cell were such as bone is discussed. homogenates Antigenic potential responses. UNITED ANTIGENS IN antigens on the cell surface. Additional studies antigens between bone cells and lymphocytes. much antigenicity ARGENTINA, procedure performed. way, providing evidence the isolation in a specific cells to clinical fields and skeletal sarcomata Although the AIRES, Abraham cells Bone CELLS CHICAGO, containing antibodies against results suggest that bone cells form, at least after cytotoxic antibodies of bone diseases bone antigens. sera RAY, Surgery, on isolated (histocompatibility) BONE BUENOS D. ROBERT From TRANSPLANTATION reprints should he addressed to Dr Niuscolo. JUL JOURNAL OF BONE ANE) JOINT SURGERY IN CELLULAR EXPERI VITRO STUDIES OF TRANSPLANTATION 343 ANTIGENS MENTS Lewis HUMORAL BN EXPERIMENTS Lews BN freeof - -.;:-:3 ___J marrow bone grafts’ <complete bone _____________ grafts E a) U.) 414. Absorption u R ES Cu LT th BN lymphoid cells 1 Lymphocyte measured by3H-Thymidine FIG. Cellular experiments Cytotoxicity Stimulation uptake following cultures served lymphocytes __;p BN cells bone cells FIG. 1 Humoral (see text). phocytes from BN rats (MLC), and 2) lymphocytes from Lewis rats + bone cells from BN rats (MLBCC). Since there are major histocompatibibity differences between Lewis and BN rats, mixed lymphocyte cultures will give rise to high stimulation. The results were compared with those obtained in mixed lymphocyte-bone cell cultures. The against as controls: lymphocytes or bone cells from the same histocompatibility differences and, therefore, stimulation); and 2) lymphocytes from Lewis cytes from BN rats + bone cells from BN rats possible inhibitory effects of bone cells over lymphocyte cultures. + 1) lymphocytes rat strain (no no anticipated rats+ lymphoto determine regular mixed Mixed lymphocyte culture (MLC) and mixed lymphocyte-bone cell culture techniques (MLBCC)-The culture procedure has been fully described elsewhere (Muscobo et a!. 1976). Blood was collected by cardiac puncture and lymphocyte suspenwere prepared by dextran sedimentation. After washing, the lymphocytes were resuspended in culture medium (supplemented MEM Hank’s base with 10 per cent heat inactivated rat serum) and a cell suspension of 2.5 x 106 x ml (85 per cent living lymphocytes) was prepared. The cells were were split into pieces in both 2 experiments and grafted (see text). into the subcutaneous tissues flanks. Humoral cytotoxicity studies-At intervals tion, serum was obtained from the immunised studied for the presence of cytotoxic after immunisaLewis rats and antibodies in the trypan blue exclusion cytotoxicity test (Tissot and Cohen 1972), using lymphocytes and isolated bone cells from BN rats as target cells. The principle of the cytotoxic test is as follows: target cells, serum and complement are incubated in vitro; if the serum contains antibodies against major histocompatibibity the target cell surface, complement is fixed and the cell dies. The percentage of killed cells is then determined antigens on by trypan viable blue cells uptake. Dead are not (Figs. Results were expressed are - stained by trypan blue, 5). as cytotoxicity - control sample 100 cells 3 to index: xlOO control sions The five Lewis (with no mean rats major cytotoxic grafted percentage with obtained bone histocompatibility from Lewis differences; with sera from or Fisher rats not included in Figure 2) tested against lymphocytes or bone cells from BN rats (4 per cent and 41 per cent cytotoxicity respectively) mixed and cultured in 0.2 millilitre microplates. After 120 served as control values. Significance levels were calculated hours of incubation at 37 degrees Celsius in humidified 5 per t test. cent CO2 air atmosphere, 1 pCi of 3H-Thymidine (Sp. ac 6#{149}7by the Student’s Ci/Mmobe) was added to each culture. Eighteen hours later, Absorption studies (Fig. 2)-Absorption is the removal of the cultures were precipitated onto glass fibre paper and the antibody from an immune serum by treatment with particulate lymphocyte proliferation assayed by determining the 3Hantigen homologous to that antibody, followed by centriThymidine uptake using liquid scintillation spectrometry. fugation and separation of the antigen-antibody complex. Varying concentrations of bone cells were used in the Serum obtained from Lewis rats grafted with bone from MLBCC, ranging from 0.4 to 1 .2 x 106 x ml. BN rats was divided in two groups: one was untreated, a The results were expressed as the mean count per minute second was absorbed with BN lymphoid cells. The cytotoxic ( ± standard Humoral error) experiments Immunisation with complete marrow (cortex VOL. 9 59-B, of four No. replicate (Fig. cultures. 2) animals-Twenty Lewis rats were grafted bone (cortex and marrow) or bone free of minus marrow) from BN rats. The bones of 3. AUGUST 1977 activity of both serum groups was then tested against cytes and bone cells from BN rats as described. The purpose of this part of the experiment determine if lymphocytes and bone cells share gens. If so, both should show none or at least cytotoxicity when tested with the absorbed sera. bymphowas surface to anti- diminished 344 D. L. MUSCOLO, S. . KAWAI AND R. D. RAY . .77 . ,)i 3 FIG. Figure 3-Isolated medium, uptake and Absorption was cell (Wright, Hargreaves, 1973) by incubation with 2 x 1O Celsius. After stored medium. performed serum and culture contrast. excluding undiluted Specificity samples with in blue. Phase lymphocytes and HeblstrOm at 37 degrees suspended of trypan two living Bernstein removed FIG. bone shows lymphocyte ;... at -20 controls lymphoid lymphoid (x the cells for Bansal, sixty Celsius until 5 FIG. (x 1200.) Figure 4-Isolated hone cell suspended in showing uptake of trypan blue in culture medium. Phase contrast. culture by one ( x 900.) dead Li of minutes the serum was used. were done by absorbing cells from “third party” 4 conrast. 1200.) Figure 5-Photomicrograph dye. The cells are suspended of2 milbilitres centrifugation, degrees Phase (I) +1 aC 8000 Li some Buffalo sera rats. Fa- Li z RESULTS Cellular in vitro Mixed major rats+ high experiments (Fig. 6) lymphocyte cultures between rats that differ at the histocompatibility locus (lymphocytes from Lewis lymphocytes from BN rats) gave, as expected, and (7898 reproducible C.P.M. lymphocyte-bone combination from BN levels ±SE cell (lymphocytes rats) did of lymphocyte On 556). the cultures from not show 000 I stimulation other with Lewis 2000 I.;- hand, mixed the same strain rats + bone cells lymphocyte stimulation in any of six consecutive experiments (49 C.P.M.±SE 7). These values were practically the same as those obtained in the control experiments lymphocytes or bone cells cultured therefore, (with with from which the same no histocompatibility no expected stimulation) This suggests isolation procedure culture conditions lymphocytes The in isolated bone some ruled inhibitory effect over lymphocyte out. Mixed lymphocyte cultures cells were 202) with from added (lymphocytes and bone lymphocyte and no significant regular mixed Lewis rats Serum raised BN differences lymphocyte + lymphocytes in Lewis may have stimulation in which was bone Lewis rats + from cells from stimulation Humoral cytotoxicity (Fig. 7 and Table I) cells rats) (7315 showed C.P.M. high ±SE were found (P >050) cultures (lymphocytes from BN rats) rats with bone grafts culture (MLC) and mixed lymphocyte-bone cell results. Cultures were harvested after 120 hours after hours pulse with 3H-Thymidine. Values for 3H-Thymidine uptake represent the lymphocyte stimulation in each culture. Note that bone cells alone were incapable of stimulating albogeneic lymphocytes in vitro. Lymph. = lymphocytes; Lewis= (MLBCC) eighteen Lewis from grafted BN rats animals) rats; BN = Brown (fifteen showed Norway rats. antisera from nine different 69 ± SE 2 cytotoxic index over BN lymphocytes and (P< 0.#{216}J1) Serum raised 37 ± SE in Lewis 4 over rats with BN bone bone grafts cells free of marrow from BN rats (twelve antisera from six different grafted animals) showed 60± SE 5 cytotoxic index over BN lymphocytes and 31 ± SE 3 over BN bone cells (P < tested from complete lymphocyte 0001). The experiments 6 FIG. Mixed culture that bone cells, at least after the that was performed and under used, did not stimulate allogeneic in vitro. possibility that lymphocytes levels of and, 49 CULTURES lymphocytes+ rat strain were differences (Fig. 6). ,,( ___________ specificity over Buffalo “third rats, of these two groups party” lymphocytes and no significant of antisera was and bone cytotoxicity cells was found. THE JOURNAL OF BONE AND JOINT SURGERY IN VITRO STUDIES OF TRANSPLANTATION These plantation 70 60 Li (I, r? rLL&I Sero ... Sero raised with bone free of marrow raised complete . . . +1 with bone grafts. toxic levels Absorption x Li 0 Absorption lymphoid z 40 carried >- I- 0 >< 0 BN 20 cytotoxic control bone, found tests (Table anlayses and bone were Bone was Cells cally (unabsorbed: values as cytotoxicity - sample - 100 It been responsible a limited x 100. control CvioToxlcrrv OF HUMORAL BONE Significance levels CELLS AND were grafts serum from after receiving BN rats INDICES OBTAINED LYMPHOCYFES calculated using bone bone on the effect from over Control sera from Lewis five different BN lymphocytes Finally, Lewis marrow the obtained Fisher rats) showed and 0±SE cytotoxic rats with BN complete bone were compared Lewis rats rats (twelve 59-8, No. 3. AUGUST 1977 grafted sera at least able the 6 and to to absorb second. transplantation antigens lymphocyte stimulation (Lane and Ling 1973), have and AGAINST target ISOLATED t test cells Bone cells *37±4 t 1 ] T I t=1#{149}20 05>P>0#{149}2 31±3 indices of serum with samples 0± SE 2 cytotoxic 3 over BN bone index cells. raised bone and BN free-of(Table I). When lympho- cytes were used as target cells, the differences of the first were significant (005 > P > 002), cells were the target, there was no significant (0.5 > P > 02) between both groups. VOL. 13, 8 and grafts from or over bone dramati- lymphoid surface, were when BN dropped that their first the in vitro distribution Also, over absorbed: indicate 60±5 bone 29; Humoral dropped to 69, 67 and taken sera with t:01 t=21 005>P>002 Free-of-marrow 49 and , t - cells. DISCUSSION proposed that Lymphocytes grafts lymphoid absorbed cytotoxic the Student 69±2w Complete two raised free-of-marrow were antigens for tissue BN Lewis same I TABLE COMPARISON the the identity of antigens were 6 respectively). results since with has the 31 share antibodies index: control using lymphocytes (unabsorbed: indices for extent, results. Sera raised with complete bone grafts bone grafts gave positive cytotoxicity against cells or lymphocytes from the donor. Cytotoxicity recorded cells transcyto- samples, with BN BN 2 and These cells some 7 with over BN absorption 3, the bone CELLS FIG. Humoral cytotoxicity or with free-of-marrow either isolated bone bone antisera and one cytotoxic respectively). TARGET over cells share but lower II) absorbed cells, ---- BN that bone lymphocytes, to determine further cell transplantation indices levels after humoral activity were 65 ; absorbed: 0 Lymphocytes indicate with out. Three different complete bone 30 0 I0 F- results antigens antisera. grafts. sera. Control 345 ANTIGENS in favour but if bone difference studies induce on the ability such stimulation (Main 1973), 1967) in and incapable brain thyroid cells of cells have cells (Lane, of stimulating other been than lymphocytes reported. Fibroblasts (Pulvertaft Jackson allogeneic have 1972), sperm chondrocytes to be good also been tested cells (Levis (Gertzbein stimulators. Ling lymphocytes On the other hand, epidermal cells Jones and Kountz 1971), endotheial Burger recently proved and and (Main, cells to Pulvertaft 1975) are in vitro. Cochrum, (Vetto and and Whalen 1976) and and Lance 1976) Neoplastic bone cells (V#{225}nky, Stjernsw#{228}rd and Nilsonne 346 D. L. MUSCOLO, S. KAWAI TABLE CvroToxIc ABsoRvrloNs REACTIONS PERFORMED WITH AND D. RAY II UNABSORBED WITH R. AND LYMPHOID ABSORBED CELLS SERA BN FROM RATS’ RN target Lymphocytes Serum Lewis rat wth grafted Unabsorbed Absorbed Lewis rat wth Unabsorbed grafted Absorbed Lewis rat grafted with BN Cytotoxic Unabsorbed 1972), results are but antigens the bone presence it difficult as cytotoxicity of the activity of tumour- to extrapolate the This observation difficulty bone should be taken possible bone cell variables concentration of obtaining cells of the per isolated determined and this more bone proved for antigen it may density no sufficient study, are (Edidin thought reported tissues that isolated cent it was to cells and, on Loghem 1966) kidney Belzer and Lance 1971), and their cell (Engelfriet, have lympho- Kountz sperm a wide chondrocytes 1976) were antibodies, antigens. Although presence of cells (Elves tissue antiin this test distribution studies have been from different solid of SD transplantation surfaces. Heersche, It was Eijsvoogel cells (Perkins, 1975), transplantation cytotoxic several endothelial cells (Vetto Gantan, epidermal found and and Siegel, cells (Fellous and 1974a; Malseed killed by, or were able showing cell membrane Langer, demonstration serological antigens cells The indices Czitrom, has with Pritzker and Gross 1975), of SD transplantation been no antigens on reported. that van Burger Howell, (Cooper and Dausset 1970), and to absorb, expression Heyner cytotoxic of those evidence in bone for the (Elves isolated bone differences cell lines both were bone of the humorab cytotoxicity for the presence of SD Sera raised in the cell-antigen were found, since significantly cells. cells. experiments transplantation higher Absorption expression cytotoxicity against studies with lymphocytes confirmed a “share” of SD transplantation antigens, since lymphoid cells were able to absorb most of the antibodies with cytotoxic effect over bone cells. The remnant cytotoxicity a that was found after absorption could the possible presence of differentiation antigens on bone cells, as described there The was no attempt expression these been so-called explosive to confirm Frederiks, HL-A due to Burwell, of transplanted Van Oud Van Bone Gowland TI-fE antigens antigens” has the increasing in close relation conditions. play a fundamental with and role Hooff, Pena OF same tissues Dexter BONE in the and AND Van between species, elicits do (Chalmers 1963). grafts with weak antigenicity 1976) seem to preserve JOURNAL a great foreign tissues Alblas, Keuning, allotransplantation, different individuals within the transplantation immunity as other freeze-dried bone 1976; Friedbaender speculation. are Termijtelen, 1975). by antigens at the cell surface is by a chromosomab region called HL-A in the human. Interest in acceptance or rejection (Van Rood, Blusse Leeuwen this stimulating “transplantation in recent years evidence that they number of pathological HL-A antigens be explained or tissue-specific for other cell lines, of lymphocyte and SD transplantation genetically determined Ag-B in the rat and 1959; there is transplantation 6 1974b; than therefore, allogeneic (SD) humoral have 6 formal Quantitative between not alive”, obtain Eijsvoogeb bone membranes using intact cells isolated to determine the presence antigens fibroblasts 1971), to 29 complete bone or with bone free of marrow were abbe to kill lymphocytes and isolated bone cells from the donor. These reactions proved to be immunologically specific, since control and “third party” reactions were negative. and to stimulate Recently, 65 index (see text). Note that absorption lymphocytes and bone cells. but 1972). 8 in the expen(due to the cytes defined by the 2 on stimulation. “strength” 49 antigens “metabolically as essential be possible in their in vitro. Serologically gens, demonstrated 67 as preliminary deleterious effect of collagenase isolate bone cells, on antigens lymphocyte Moreover, 13 provide alive, (Schellekens 1970). 3) The possible and EDTA, used to 3 results evidence million cells in 60 to 70 per to be they were reported stimulation responsible one 2) Although whether has been lymphocyte than milbilitre). cells for bone cells. due to the following ment: 1) Inadequate . recorded most The present studies suggest that isolated bone incapable of stimulating allogeneic lymphocytes vitro. low probable makes to normal Absorbed activity was cells removes lymphoid ; Han 31 bone free-of-marrow 1971 69 BN complete bone associated Bone cells BN complete bone S cells Even (Burwell transplanta- JOINT SURGERY IN tion antigens (Urist, abbe to Mikulski persistence generate and of those VITRO an Boyd immune 1975), antigens STUDIES OF cells due gens 1975). In addition duction of lymphocytes, to being “destructive” HL-A circumstances so far unclear, antibody called the graft (Carpenter, experimental since they profile of bone cells, system may provide problem. Association of diseases antigens and psoriasis reported pro- HL-A antigens, supporting and certain genetic origin of these HL-A B27 with Reiter’s established infections of to protect Salmonella arthritis and interactions better with HL-A antigens with Several reported diseases of bone to be associated HL-A B27 with spondylitis well accepted. This antigen of patients Fries this occulta Gallmeier, mas, leukaemias, spondylitis abnormal also be and developments included in this asymmetry of the of the group. facet 1975), Terasaki, theory (Takasugi, Terasaki, 1973; and and and Delbon lympho- solid tumours Mickey, Menck Chretien, 1975) with specific at present BOhme, 1972), other Henderson, have and Rogentine, showed significant HL-A antigens. it is unclear how transplanta- relate to the resistance or susceptiauto-immune and infectious diseases transplantation throw light suggest antigens present induced sarcomas are modified or altered antigens present in normal cells Baldwin 1975 ; Invernizzi and Parmiani Better charactensation histocompatibility into the malignant been of after or rheumatoid Stiehm 1974) Kuwert, Schmidt Tarpley, tumour-associated 1975). Voak of an immuno- and juvenile Katz and (Bertrams, sarcomas in chemically histocompatibility (Bowen and (Calm have the Wetter Thompson that lumbo-sacral Spina bifida joints and in man, future developments in this field may on the aetiology of these conditions. Finally, recently presented observations and probably association of disorders is now in approximately ankylosing Hazleman reported. tion HL-A antigens bility to neoplastic, 1975). Some spine may Reis, antigens 1975). (Russel, Shultes and Kuban in association with particular myeloma associations Although and joint have with increased and related is present with particular of transplantation from the normal of a specific HL-A antigen, be in the future. The strong cent immune of been Twomey been frequency more will the also HL-A Johnson conditions. The association disease and acute arthritis with Shigebla, Yersinia (Brewerton (Rachelefsky, Multiple et a!. 1975) in ofthe antigenic understanding would imply distribution in those patients different population. recently have specific and (Bulgen, and arthritic have been of a type Jeter 1972; Langer Further definition shoulder 1976) 1972) the tend with Mendell anti(Elves from immune recognition or destruction D’Apice and Abbas 1976). There is evidence suggesting that this may be a (Bonfiglio transplantation. 95 per for antibodies under the production “blocking”, factor bone and responsible cytotoxic antigens trigger, association Ruderman, Frozen or cell debris. However, the role of cellular transplantation in the fate of bone grafts is far from clear in (Amos, to 347 ANTIGENS reported response possibly on dead TRANSPLANTATION of antigens may transformation cell surface provide further of the cells. bone insight REFERENCES Amos, R., Mendell, D. B., Proceedings, Ruderman, Bach, F. H., activation. Bach, M. L., and Nature (London), Bard, D. R., Dickens, Journal of Bone 7, Supplement N. R., and Johnson, H. (1975) Linkage between HL-A and spinal development. Transplantation 1, 93-95. Sondel, P. M. (1976) 259, 273-281. M. J., Edwards, .J., and Smith, and A. Joint Surgery, 56-B, Differential function A. U. (1974) Studies of major on slices histocompatibility and isolated complex cells from fresh antigens in T-lymphocyte osteoarthritic human bone. 340-351. Bertrams, J., Kuwert, E., Bohme, U., Reis, H. E., Gailmeler, W. M., Wetter, 0., and Schmidt, C. G. (1972) HL-A antigens in Hodgkin’s disease and multiple myeloma. Tissue Antigens, 2, 41-46. Bonfiglio, M., and Jeter, W. S. (1972) Immunological responses to bone. Clinical Orthopaedics and Related Research, 87, 19-27. Bowen, J. G., and Baldwin, R. W. (1975) Tumour-specific antigen related to rat histocompatibility antigens. Nature (London), 258, 75-76. Brewerton, D. A. (1975) HL-A 27 and disease. Journal of Bone and Joint Surgery, 57-B, 247. Bulgen, D. Y., Hazleman, Burwell, R. C. (1976) Burwell, R. G., Gowland, A., Journal and Chalmers, Edidin, M. Kahan, Elves, B., D’Apice, in Immunology, 22, F., and 1-65. E. M. M. W. (1974b) M. W. (1975) A study C. P., Heersche, by means No. of cytotoxic 3. AUGUST immune of the response J. N. M., ELjsvoogel, antibody test. Vox 1977 in bone (1976) frozen shoulder. The for detecting locations Academic antigens to allografts of allogeneic V. P., and sanguinis, spondylitis homografting. method transplantation behaviour ankylosing A. K. and cellular New York: A., and HL-A-B27 Lancet, 1, 1042-1044. allografts. Transplantatiofi Proceedings, 8, Supplement 1, 95-111. Studies in the transplantation ofbone. VI. Further observations concerning Journal of Bone and Joint Surgery, 45-B, 597-608. of A serological of the Humoral Studies Abbas, immunity (1971) (1972) The tissue distribution B. D., and Reisfeld, R. Elves, 59-B, J. A. Transplantation Lance, Elves, VOL. (1976) bone prevalence M. W. (1974a) 56-B, 178-185. Engelfriet, D. Fries, J. F. (1975) Striking of Medicine, 293, 835-839. and G., cortical J. (1959) S., Voak, F. (1963) bone. Carpenter, C. Advances Cooper, and fate of freeze-dried and Dexter, and cancellous of homologous Calm, B. L., The role of antibodies Journal the of Bone van Loghem, 11, 625-630. and antigens antigens. on chondrocytes ofbone. in the surface of transplantation Press. cancellous W27 in “healthy” from rejection Joint In articular J. J. grafts (1966) in inbred Demonstration males and 41-B, cells. Transplantation cartilage. Journal andApplied rats. Transplantation, ofleucocyte and females. enhancement Surgery, ofepidermal InternationalArchivesofAllergy bone positive the antigenicity New of organ England allografts. 160-179. Transplantation, Antigens, of Bone 11, 108-109. pp. 125-140. and Joint Ed. Surgery, 47, 708-715. Immunology, 19, 416-423. iso-antigens on skin fibroblasts 348 D. L. MUSCOLO, 5. KAWAI AND R. D. RAY Fellous, M., and Dausset, J. (1970) Probable heploid expression of HL-A antigens on human spermatozoon. Nature (London), 225, 191-193. Friedlaender, G. E. (1976) The antigenicity of preserved allografts. Transplantation Proceedings, 8, Supplement 1, 195-200. Gertzbein, S. D., and Lance, E. M. (1976) The stimulation of lymphocytes by chondrocytes in mixed cultures. Clinical and Experimental Immunology, 24, 102-109. Han, 1. (1972) Blastogenic response of normal lymphocytes to cultured lymphoid cells and non-lymphoid neoplastic cells. immunology, 23, 355-359. Invernizzi, G., and allogeneic Lane, J. T., 250-259. Lane, Jackson, T., j. and various Langer, F., McDevitt, Main, S., Schellekeus, culture. and M., Cancer Urist, R., Mikulski, cells. induced A function syngeneic effects offresh leukocytes response genes, and sarcomata cross of maturation. in mixed frozen of the Cellular as a measure and Transplantation, profile and with Transplantation, cultures allogeneic reacting with bone. tissue 19, cells from JournalofBone and disease. of histocompatibility Lancet, in man. Science, 191, 1, 1269-1275. 15, 247. DNA synthesis ofthe U.S.A., rat chondrocyte. Arthritis and humoral immune response bone Clinical Orthopaedics Kuban, Beizer, Tissue Activation and D. J. antigens F. 0., and in mixed cultures 68, 1165-1168. of rat and 19, 223-23 Rheumatism, analysis leukocytes and allogeneic 1. of bone-allografted rats. Journal of Bone (1972) E. R. S. L. (1974) Histocompatibility 7, 229-239. B., Mickey, M. G. N., Jun., Twomey, R., (1975) Journal of Clinkal Increased prevalence (HL-A) transformation immunology, and Related 87, 156-164. Research, 11, 175-183. Transplantation, Reactions of kidney cells-with cytotoxic antisera: 5, 88-98. of lymphocytes. Stiehm, in the rat. Kountz, Antigens, V. P. (1970)Lymphocyte Henderson, grafts. histocompatibility Howell, E., antigens. Experimental P. I., osteo-articular of Ag-B Katz, R., 290, 892-893. Etjsvoogel, Meuck, and III. 20, of W27 antigens in vitro. H., Pathology, associated with Mechanism W. 795-805. in juvenile rheumatoid psoriasis. New of stimulation Thompson, R. (1973) A. L. (1975) Histocompatibility arthritis. England in the HL-A mixed antigens New Journal lymphocyte in solid tumors. 648-650. P. B., Rogentine, ofSurgery, R. G., and Cohen, M. and I. (1967) P. I., 33, Tarpley, J. L., Chretien, neoplasms. Archtves Tissot, osteo Pulvertaft, Terasaid, and and Kountz, S. L. (1971) National Academy ofSciences Antigenic Z., Siegel, S., kidney-specific and and immune R. D. (1976) analysis L. M., 738-740. Research, and immunogenicity of sperm et al. (letter). ofthe of Medicine, P. T. A., Clinical skin of chemically 826-832. Terasaki, T. J., Shultes, of Medicine, 287, with response A. E. (1975)The cultures M. J., (1976) Massive for Journal antigens stimulation allogeneic HL-A, Mardiney Ray, Immunogenetic G. S., England S. and 58-A, R. J. V., R.achekfsky, Lymphocyte Gross, F. (974) to Proceedings H. A., Gantan, Pulvertaft, and Mixed K. C., Jones, cells. C. E. (1972) Takasugi, W. reference evidence (1975) Rat thymocytes-the 16, 602-609. J. J. (1976) M., and Heyner, J. (1971) R. K. P., Bodmer, In skin possible Russel, and D. L., Kawal, Joint Surgery, Ottolenghi, N. transplantation 254, 713-714. Tumour-associated Nature (London), 216-220. Cochrum, Z. Perkins, Ling, Pritzker, Whalen, (1973) R. K., dissociated Palm, A., H. O. Muscolo, and (1975) antigens. and 57-A, and R. K. Malseed, G. N. R. (1973) Transplantation, Ling, Czitrom, Surgery, W. R., 302-304. Main, L., organs. Joint Levis, Parmiani, histocompatibility C. (1972) A., Histocompatibility Boyd, and Surgery, 110, 416-428. V#{225}nky,F., Stjernsward, J., and P. L., and Dellon, antigens and solid malignant 110, 269-271. in the rabbit. S. D. (1975) A U. NIISOnne, Identification (1971) Cellular of the major antigen-extracted chemosterilized immunity to human autodigested sarcoma. locus. Tissue alloimplant Journal of the Antigens, 2, 267-279. for bone National banks. Archives Cancer of 46, Institute, 1145-1151. Van Rood, (1975) Vetto, R. J. J., Blusse Van Oud Histocompatibility M., and Burger, Aiblas, A., Keuning, and transplantation genes D. R. (1971) The identification lymphocytes. Transplantation, 11, 374-377. Vetto, R. M., and Burger, D. R. (1972) Endothelial Wright, P. W., Hargreaves, R. E., Bansal, S. C., serum factors Proceedings from ofthe tolerant National animals Academy 3. J., Frederiks, antigens. cell Bernstein, and stimulation I. D., E., Termljtelen, Transplantation comparison of transplantation of allogeneic and HelistrOm, block lymphocyte-mediated ofSciences ofthe U.S.A., 70, 2539-2543. that A.,Van Proceedings, Hooff, J. P., Pens, antigens lymphocytes. on canine Transplantation, K. E. (1973) immunity in THE A. S., and 1 , 25-30. 7, Supplement Allograft vitro are JOURNAL vascular Van Leeuwen, endothelium A. and 14, 652-654. tolerance: presumptive soluble antlgen-antibody OF BONE AND evidence that complexes. JOINT SURGERY