Europ. J. Canter Vol. 13, pp. 355--359. Pergamon Press 1977. Printed in Great Britain
T and B Lymphocytes and Monocytes in the Spleen in Hodgkin's Disease: the increase in T Lymphocytes in Involw:d Spleens A. SANTORO,* B. CAILLOU** and D. BELPOMMEt:~ *Instituto di Patologia Speciale Medica dell'Universita degli Studi di Messina, Messina, Italy ~flnstitut de Gandrologie et d'Immunogg,nttique, H6pital Paul Brousse, 14 avenue Paul Vaillant-Couturier, ViUejuif, France "~Institut Gustave.Roussy-94800 Villejuif, France, 16 his, avenue Paul- VaiUant-Couturier, Villejuif, France Abstract---The percentages of T and B lymphoqytes in the spleens of 40 patients suffering from Hodgkin's disease were studied. The results are summarized as follows: in involved spleens, i.e. containing Reed-Sternberg cells (RSC), there is significantly more T lymphocytes associated with significantly less B lymphocytes, in comparison to non-involved spleens. No significant difference was found between the two groups in the percentage of EA and EAC rosette-forming cells and non-T non-B mononuclear cells. There was no relationship between the histological type of the spleen and the percentage of either T or B lymphoqytes. The demonstration of a higher percentage of T lymphocytes in 14 spleens containing detectable Reed--Sternberg cells (RSC) out of the 40 studied confirmed our preliminary data suggesting the existence of a local immunological reaction against ReedSternberg cells, in involved spleens in Hodgkin' s disease.
INTRODUCTION
and histological criteria, and staging conformed to the Ann Arbor classification. Before laparotomy, histological examination of pathological lymph nodes showed one patient to have lymphocyte predominance (LP), 21 to have nodular sclerosis (NS) and 11 to have mixed cellularity (MC). There were no cases of lymphocyte depletion (LD); 7 cases showed typical Reed-Sternberg cells (RSC) but could not be histologically subtyped for technical reasons (ND). Clinically, 2 patients were at stage I, 19 at stage II, 16 at stage IlI and 3 at stage IV. After staging laparotomy, histological examination of the 14 spleens containing RSC (i.e. involved spleens) showed 6 NS varieties, 5 MC and 3 ND. Only 3 of the 21 patients at clinicals stages I and II had involved spleens whereas 11 out of 19 spleens were involved in the cases at disseminated stages III and IV (0.01 > P > 0.001). Spleen weight was evaluated in each case. In our series, spleen weight was 309"5 + 145 (mean value) for involved spleens, and 176 + 87
Dtrm~o the past few years several methods of investigation in man have permitted the identification of T and B lymphocytes [1-14] and of a third mononuclear cells population including monocytes [6, 15]. With different immunological techniques it is possible to characterize the mononuclear cells populations by different cell membrane markers (Table 1). The purpose of this paper is to report our findings on the application of these techniques to the study ofT and B lymphocytes and monocytes in the spleens of patients with Hodgkin's disease.
MATERIAL AND METHODS 1. Patients The spleens of 40 patients with Hodgkin's disease (17 women and 23 men, aged from 11 to 61 years) were investigated. Hodgkin's disease was diagnosed on clinical 355
356
A. Santoro, B. CaiUou and D. Belpomme Table 1. Immunological test procedures used to differentiate T and B lymphocytes and monocytes
Immunological test E Rosettes* (SRBC receptors) EA Rosettest (Ig receptors) EAC Rosettes++ (Activated C'3 receptors) Membrane Ig §
T lymphocytcs
B lymphocytes
Third categories¶ Monocytcs and histiocytcs
+
0
0
(+ )
(+)
(+ )
0 0
(+ ) +
(+ ) 0
*Spontaneous rosettes with SRBC (see text). tRosettes with sensitized SRBC (see text), ~Rosettes with SRBC antibody complement complexes (see text). §Membrane immunoglobulins detected by direct immunofluorescence. ~Non T non B mononuclear cells (see text). (+) On some mononuclear cells.
(mean value) for non-involved spleens. This difference is significant (P < 0'001).
2. Mononudear spleen cell purification The spleen fragments were gently disrupted and the cell suspension washed in Hank's medium. The mononuclear cells were isolated by lowering the suspension on a ficoll-triosil density gradient, according to a technique previously described [1-3]. The purity of the mononuclear cells was always checked by May-Grunwald-Giemsa staining and their viability determined by the try9an blue dye exclusion test. 3. Immunological tests procedures for T and B cell quantification 3.1. T l)~phocytes. T lymphocytes were enumerated by the E rosette test using sheep red blood cells (SRBC) according to a technique already described [1-3]. Rosettes were defined as lymphocytes surrounded by at least 3 SRBC. 3.2. B lymphocytes. B lymphocytes were enumerated by determining the membrane immunoglobulins (mIg) by a direct immunofluorescent test using a polyvalent fluorescent isothiocyanate conjugated sheep anti-human immunoglobulin serum, as previously reported [1-3]. 3.3 Ig receptor (EA) and activated complement receptor (EAC) bearing mononuclear cells. Enumeration of these mononuclear cell categories was performed by the EA and EAC rosette tests respectively. Details of these tests have also been reported elsewhere [1-3]. 3.4. Non- T non-B mononuclear cells (n T nBmc). In addition, nT nBmc were calculated from the
following formula: nT nBmc = m c - ( T + B ) , in which mc = total number of mononuclear ceils, T = total number of T lymphocytes, B -- total number orB lymphocytes [16].
4. Controls and statistical analysis Peripheral blood lymphoeytes from 20 normal subjects were used as control and the results provided the normal range for each test. Statistical analysis were performed using the Student t-test. RESULTS
(a) Immunological findings The results are summarized in Table 2. In non-involved spleens (absence of detectable RSC), the mean percentage of E rosettes (T lymphocytes) was 35"5%. Extreme values ranged from 15 to 67%, with only 3 cases above 50%, whereas in involved spleens (presence of RSC), the mean percentage of mononuclear cells forming spontaneous E rosettes (T lymphocytes) was 50%, with extreme values ranging from 33 to 67% and only 3 cases below 42%. This difference is significant (P < 0.01). For EA and EAC rosettes, mean percentages were 16.5 and 22% respectively, for non-involved spleens. Extreme values ranged from 3 to 33% for EA rosettes and from 1 to 65% for EAC rosettes. In involved spleens, the mean percentages of EA and EAC rosettes were 17 and 14.5% respectively. Extreme values ranged from 1 to 51% for EA rosettes and from 1 to 35 ~ for EAC. This difference is not significant. In non-involved spleens, 31.5 % of the mono-
T and B L ymt~cytes and Mo~ytes in the Spleen in Itodgkin's Disease
357
Table 2. Membranemarkersin mononudearspleencells (Hodgkin'sdisease) B lymphocytes (membrane Ig) %
Conditions
No.
Non-involved
Mean Extreme value values 33.5*
26 Spleens
14 "Spleens
(E rosettes)
EA Rosettes
EAC Rosettes
M e a n Extreme value values
M e a n Extreme value values
M e a n Extreme value values
33.5t
_+ 11.5
ll6S
Extreme values
31(+ 16) 0165
14.5
1/51 4- 14
Mean value
=k16
17
33167
Non-T non-B mononuclear cells %
22
3/33 + 12
50 t
7140 _+ I I
16.5
15167 + 13.5
23*
Involved
T lymphocytes
13170 + 13
E A and E A C binding mononuclear ceils
1/35 27(+ 13) 8/40 :i: 12
*P < 0.05 t P < 0.01.
nuclear cells (ranging from 13 to 70%) showed surface bound Ig (B lymphocytes). In involved spleens, the mean percentage ofsplenic lymphocytes bearing Ig on the cell surface was 23%, with extreme values ranging from 7 to 40%. This difference is significant (P < 0.05). Table 2 also shows the percentages of non-T non-B mononuclear cells (nT nBmc). In noninvolved spleens the percentage was 31% (range: 0-63%) and in involved spleens 27% (range: 8-40%). No significant difference was detected. (B) Searchfor a correlation between immunological data and variables characterizing the disease This search led to three interesting observations: (1) We found a significantly higher percentage of splenic T lymphocytes in disseminated clinical stages III and IV than in cases of localized disease (stages I and II). We also found a slightly fewer mIg binding mononuclear cells, (B lymphocytes) EAC rosetteforming cells and nT n.Bmc in the generalized disease. However, these differences were not significant. No variation was detected in the percentage of splenic EA rosette-formlng cells in patients at different clinical stages of the disease. (2) We compared involved spleens in the localized and disseminated stages and found no difference in the percentages of T lymphocytes. Nor was any variation detected when comparing T cells of the non-involved spleens in each clinical group. (3) Finally, we found no clear-cut relationship in the spleen between the percentages of
the different mononuclear cell populations and the histological type of the tissue. DISCUSSION The results presented in this paper confirm our previous findings [17-21] as well as those of others [22, 23] that in Hodgkin's disease there is a significantly higher percentage of T lymphocytes present in spleens containing RSC, than in non-involved spleens. The mechanism of this increase is at present unclear. There are at least two possible explanations: (1) Since it has been claimed that RSC may be T cell derived, this increase in T mononuclear cells may correspond to an increase in RSC precursors. This hypothesis does not seem plausible, however, because recent reports, including our own [18-21], have suggested that RSC may in fact, be derived from B cells [24, 25] or from histiocytes [26, 27]. (2) This increase in T cells may correspond to a local immune reaction against RSC. This would explain the strikingly close contact seen with conventional [28] and scanning electron microscopy [18--20] between presumed T lymphocytes and RSC. These morphological findings do not, however, prove the existence of a specific immunological reaction. Our data regarding B lymphocytes confirm reports from other authors [29, 30] who found in using different methods a significant drop in the number of B cells in involved spleens. However, from our data we are unable to
358
A. Santoro, B. Caillou and D. Belpomme
deduce that there is a decrease in the absolute number of B lymphocytes in involved spleens in Hodgkin's disease. The absence of any significantdifference in the percentages of E A and E A C rosette-forming cells suggests that there is no difference in the percentage of cells related to the histiocytic ScI~eS. We showed a higher mean percentage of splenic E rosettes (T lymphocytcs) in disscminated clinical stages Ill and IV than in localized stages I and II. According to our findings,there isno differencein the percentage of E rosette forming cells between involved spleens, whether they arc localized or disscminatcd diseases. Our data showing a high mean percentage of splenic T lymphocytcs in the generalized clinical stages may in fact bc
explained by a larger number of involved spleens in the disseminated cases. There is however, no clear correlation between the increase of T lymphocytes in the spleen and the degree of dissemination of the disease. Our findings suggest that there is no correlation between the percentage of the different mononuclear cells and the histological type in the spleen; however, as our series was exclusively composed of NS and MC varieties, no definite conclusions can be drawn from these data. Finally, from this study it is suggested that the detection of an increased number of T lymphocytes in involved spleens may be clinically helpful in the evaluation of the clinical stage of Hodgkin's disease when pathological analysis remains difficult to interpret.
REFERENCES
D. BELPO~, D. DA~rc~mv, E. Du Rus~Lum, D. GRANDJON,R. HUCHET, P. POUILLART,L. SCHWA~ZE~B~RG,J. L. A~m~L and G. IVIATF~, T and B lymphocyte markers on the neoplastic cell of 20 patients with acute and l0 patients with chronic lymphoid leukaemia. Biomedicine, 20D 109 (1974). 2. D. BELPOM:~S,D. DA~TCH~V, E. Du Rus~uEc, D. GRA~rDJON,R. Huc~mT, F. PrNoN, P. POLULLA~T,L. SC~WARZENBEXO,J. L. A~EL and G. MATm~, La nature T ou B des cellules ndoplasiques des leuc&nies lymphoides. Bull. Cancer61~ 387 (1974). 3. D. BEx.POm~m, D. D~'~TCm~V, R. JosEPH, R. Hucn~% A. Sa_wroa~o, D. G~joN and G. M~TH~, Further studies of acute and chronic leukemias: T and B cell membrane markers and scanning electron microscopy. In Current Studies on Standardization. Problems in Clinical Pathology, Haematology and Radiotherapy in Hodgkin's Disease (Edited by G. ASTAnDI, C. B~on~I, M. ~sA, L. T~lcro~ and G. TORLO~,'rANo)p. 143. Excerpta Medica, Amsterdam (1975). 4. C. BL~co, R. PATmClCand V. NUSSENZWEm,A population of lymphocytes bearing a membrane receptor for antigen-antibody complement complexes. J. exp. Med. 132, 702 (1970). 5. W . H . LAY, N. F. MEND~S, C. R. Bunco and V. Nuss~.NzwmG, Binding of sheep red blood cells to a large population of human 1ymphocytes. Nature (Z~nd.) 230, 531 (1971). 6. W . H . LAY and V. NussENzvrzm, Receptors for complement on leucocytes. J. exp. Med. 198, 991 (1968). 7. G. ~'IICHLMAYRand H. Hus~R, Receptor sites for complement on certain human peripheral blood lymphocytes. J. Iramunol. 105, 670 (1970). 8. M. PAPAmCHPaL, E. J. HOLBOROW, H. I. K~rrH and H. K. F. CuR~x,, Subpopulatlons of human peripheral blood lymphocytes distinguished by combined rosette formation and membrane immunofluorescence. Lancet 11, 64 (1972). 9. B. Psgms, M. Fsggxaum, L. FORm and L. A~A.,~, Immunoglobulins on lymphocyte membrane. In Progress in Immunology, p. 95 (Edited by B. AMOS) Academic Press,New York (197I). 10. S. PrNcus, C. Bn~a~co and V. NusszNzwmo, Increased proportion of complement receptor lymphocytcs (CRL) in peripheral blood of patients with chronic lymphatic Icukacmia (CLL). Fed. Pro¢.31~ 775 (1972). l I. E. R~ZLLrNO, S. COLON, H. M. GmzY and E. R. UNAmy~, Immunoglobulins oflymphocytes I, Distributionand quantitation.J. exp. Med. 133~ 156 (1971). 12. E. Sh'~VACH, E. JAyFz and I. GREEN, Receptor for complement and immunoglobulinon human and animal lymphoid cells.Transplant.Rev. 16~ 3 (1973). 1.
T and B Lymphocytes and Monocytes in the Spleen in Hodgkin's Disease 13. 14. 15. 16. 17. 18. 19.
20.
21. 22. 23. 24. 25. 26. 27. 28. 29. 30.
J . D . Wn.~oN and G. I. V. Noss~, Identification ofhuman T and B lympho.. cytes in no ~rm~! peripheral blood in chronic lymphocytic leukemia. Lancet ii, 788 (1971). J. WYB~'% M. C. C~aua and H. H. FtmENBEaO,The human rosette forming cell as a marker of a population of thymus derived cells. J. din. Invest. 51, 2537 (1972). H. HtmER and H. H. Ftmm,rsERo, The interaction of monocytes and macrophages with immunoglobulim and complement. Set H ~ t . 3, 160 (1970). R.. Josm, H, D. BELPOm~ and G. MATm~, Increase in "null" cells in acute lymphocytic leukaemia in remission on long term immunotherapy, Brit. J. Cancer 33, 567 (1976). D. BELPOMME,I~. JOSEPH, L. N~VAP.ZS,R. G ~ u - M . ~ , R. HUCI-IET, I. BoT'ro, D. GRA.~JON and G. MATH~, T-lymphocytes and Keed-Sternberg cells in spleen of Hodgkin's disease. New Engl. J. Med. 291~ 1417 (1974). D. BELPOMME, D. DA~rrCHEV, R. JOSEPH, A. SANTOnO, D. GRANDJONand G. R/IATH~,CcU-membrane markers of T and B lymphocytes and monocytes in leukaemiasand hacmatosarcomas.ExcerptaMcdica, Amsterdam (1976). D. BELPO~IE, D. DA_m'CHEV, R. JOSEPH, A. SAN'roRo, N. LELARO~, D. GRANDJON, D. Po~cr-v~itT and G. MATI-~, Classification of leukaemias and haematosarcomas based on cell membrane markers and scanning electron microscopy. To be published. D. DAm'cm~v and D. BrLPO~tm~, Immunological studies of Keed-Sternberg cell and lymphocytes in Hodgkin's disease. In Proceedings of the 15th Congress of the International Society of Hematology. Jerusalem, Sept. 1974 (abstract). K.R. JOSEPH and D. B~LPOm~m,T and B lymphocytes in spleen in Hodgkin's disease. Lancet i, 747 (1975). V. Gan~o~u, G. S. Dr L GtACO, P. E. MANCO~, S. TOGN~Lt~ and G. ~ o v A x ~ , Lymphocytes in spleen in Hodgkin's disease. Lancet 332 (1975). J. KAua, A. S. D. SPIEm, D. CATOVSKVand D. A. G. G,~.m'ON, Increase of T lymphocytes in the spleen in Hodgkin's disease. Lancet il, 800 (1974). J. LEECH,Immunoglobulin positive Keed-Sternberg cells in Hodgkin's disease. Lancet ii, 265 (1973). B.H. TINI~LE,J. W. PAm~R and R. J. LUKES,Reed-Sternberg ceils in infectious mononucleosis. Amer. J. Clin. Path. 58, 607 (1972). M . M . B . KAY and M. K.~tN, Surface characteristic of Hodgkin's cells. Lancet i, 748 (1975). H. KAPPAI'ORT,Tumours of the hematopoietic system. In Atlas of Tumour Pathology, Vol. 1, Armed Forces Institute of Pathology, Washington (1966). R.B. ARCHnaAI.Dand J. F. Fm~NSaXR,Quantitative ultrastructural analysis of in vivo lymphocyte Reed-Sternberg cell interactions in Hodgkin's disease. Nat. Cancer Inst. Monogr. 36, 239 (1973). J. DIEBOLD,J. P. C~tmLE~ and G. TPacoT, Les ceilules immuno-secr~trices des rates pr~levfies au cours de laparotomies pour maladie de Hodgkin. Path. Biol. 9, 1015 (1973). R . L . LONGMIRE,R. MCI~LL, R.. YELE NOSKY, S. ARMSTRONG,J. E. LANG and C. G. CRADDOCX,In vitro splenic IgG synthesis in Hodgkin's disease. New Engl. J. ~red. 289. 763 (1973).
359
T and B Lymphocytes and Monocytes in the Spleen in Hodgkin's Disease: the increase in T Lymphocytes in Involw:d Spleens A. SANTORO,* B. CAILLOU** and D. BELPOMMEt:~ *Instituto di Patologia Speciale Medica dell'Universita degli Studi di Messina, Messina, Italy ~flnstitut de Gandrologie et d'Immunogg,nttique, H6pital Paul Brousse, 14 avenue Paul Vaillant-Couturier, ViUejuif, France "~Institut Gustave.Roussy-94800 Villejuif, France, 16 his, avenue Paul- VaiUant-Couturier, Villejuif, France Abstract---The percentages of T and B lymphoqytes in the spleens of 40 patients suffering from Hodgkin's disease were studied. The results are summarized as follows: in involved spleens, i.e. containing Reed-Sternberg cells (RSC), there is significantly more T lymphocytes associated with significantly less B lymphocytes, in comparison to non-involved spleens. No significant difference was found between the two groups in the percentage of EA and EAC rosette-forming cells and non-T non-B mononuclear cells. There was no relationship between the histological type of the spleen and the percentage of either T or B lymphoqytes. The demonstration of a higher percentage of T lymphocytes in 14 spleens containing detectable Reed--Sternberg cells (RSC) out of the 40 studied confirmed our preliminary data suggesting the existence of a local immunological reaction against ReedSternberg cells, in involved spleens in Hodgkin' s disease.
INTRODUCTION
and histological criteria, and staging conformed to the Ann Arbor classification. Before laparotomy, histological examination of pathological lymph nodes showed one patient to have lymphocyte predominance (LP), 21 to have nodular sclerosis (NS) and 11 to have mixed cellularity (MC). There were no cases of lymphocyte depletion (LD); 7 cases showed typical Reed-Sternberg cells (RSC) but could not be histologically subtyped for technical reasons (ND). Clinically, 2 patients were at stage I, 19 at stage II, 16 at stage IlI and 3 at stage IV. After staging laparotomy, histological examination of the 14 spleens containing RSC (i.e. involved spleens) showed 6 NS varieties, 5 MC and 3 ND. Only 3 of the 21 patients at clinicals stages I and II had involved spleens whereas 11 out of 19 spleens were involved in the cases at disseminated stages III and IV (0.01 > P > 0.001). Spleen weight was evaluated in each case. In our series, spleen weight was 309"5 + 145 (mean value) for involved spleens, and 176 + 87
Dtrm~o the past few years several methods of investigation in man have permitted the identification of T and B lymphocytes [1-14] and of a third mononuclear cells population including monocytes [6, 15]. With different immunological techniques it is possible to characterize the mononuclear cells populations by different cell membrane markers (Table 1). The purpose of this paper is to report our findings on the application of these techniques to the study ofT and B lymphocytes and monocytes in the spleens of patients with Hodgkin's disease.
MATERIAL AND METHODS 1. Patients The spleens of 40 patients with Hodgkin's disease (17 women and 23 men, aged from 11 to 61 years) were investigated. Hodgkin's disease was diagnosed on clinical 355
356
A. Santoro, B. CaiUou and D. Belpomme Table 1. Immunological test procedures used to differentiate T and B lymphocytes and monocytes
Immunological test E Rosettes* (SRBC receptors) EA Rosettest (Ig receptors) EAC Rosettes++ (Activated C'3 receptors) Membrane Ig §
T lymphocytcs
B lymphocytes
Third categories¶ Monocytcs and histiocytcs
+
0
0
(+ )
(+)
(+ )
0 0
(+ ) +
(+ ) 0
*Spontaneous rosettes with SRBC (see text). tRosettes with sensitized SRBC (see text), ~Rosettes with SRBC antibody complement complexes (see text). §Membrane immunoglobulins detected by direct immunofluorescence. ~Non T non B mononuclear cells (see text). (+) On some mononuclear cells.
(mean value) for non-involved spleens. This difference is significant (P < 0'001).
2. Mononudear spleen cell purification The spleen fragments were gently disrupted and the cell suspension washed in Hank's medium. The mononuclear cells were isolated by lowering the suspension on a ficoll-triosil density gradient, according to a technique previously described [1-3]. The purity of the mononuclear cells was always checked by May-Grunwald-Giemsa staining and their viability determined by the try9an blue dye exclusion test. 3. Immunological tests procedures for T and B cell quantification 3.1. T l)~phocytes. T lymphocytes were enumerated by the E rosette test using sheep red blood cells (SRBC) according to a technique already described [1-3]. Rosettes were defined as lymphocytes surrounded by at least 3 SRBC. 3.2. B lymphocytes. B lymphocytes were enumerated by determining the membrane immunoglobulins (mIg) by a direct immunofluorescent test using a polyvalent fluorescent isothiocyanate conjugated sheep anti-human immunoglobulin serum, as previously reported [1-3]. 3.3 Ig receptor (EA) and activated complement receptor (EAC) bearing mononuclear cells. Enumeration of these mononuclear cell categories was performed by the EA and EAC rosette tests respectively. Details of these tests have also been reported elsewhere [1-3]. 3.4. Non- T non-B mononuclear cells (n T nBmc). In addition, nT nBmc were calculated from the
following formula: nT nBmc = m c - ( T + B ) , in which mc = total number of mononuclear ceils, T = total number of T lymphocytes, B -- total number orB lymphocytes [16].
4. Controls and statistical analysis Peripheral blood lymphoeytes from 20 normal subjects were used as control and the results provided the normal range for each test. Statistical analysis were performed using the Student t-test. RESULTS
(a) Immunological findings The results are summarized in Table 2. In non-involved spleens (absence of detectable RSC), the mean percentage of E rosettes (T lymphocytes) was 35"5%. Extreme values ranged from 15 to 67%, with only 3 cases above 50%, whereas in involved spleens (presence of RSC), the mean percentage of mononuclear cells forming spontaneous E rosettes (T lymphocytes) was 50%, with extreme values ranging from 33 to 67% and only 3 cases below 42%. This difference is significant (P < 0.01). For EA and EAC rosettes, mean percentages were 16.5 and 22% respectively, for non-involved spleens. Extreme values ranged from 3 to 33% for EA rosettes and from 1 to 65% for EAC rosettes. In involved spleens, the mean percentages of EA and EAC rosettes were 17 and 14.5% respectively. Extreme values ranged from 1 to 51% for EA rosettes and from 1 to 35 ~ for EAC. This difference is not significant. In non-involved spleens, 31.5 % of the mono-
T and B L ymt~cytes and Mo~ytes in the Spleen in Itodgkin's Disease
357
Table 2. Membranemarkersin mononudearspleencells (Hodgkin'sdisease) B lymphocytes (membrane Ig) %
Conditions
No.
Non-involved
Mean Extreme value values 33.5*
26 Spleens
14 "Spleens
(E rosettes)
EA Rosettes
EAC Rosettes
M e a n Extreme value values
M e a n Extreme value values
M e a n Extreme value values
33.5t
_+ 11.5
ll6S
Extreme values
31(+ 16) 0165
14.5
1/51 4- 14
Mean value
=k16
17
33167
Non-T non-B mononuclear cells %
22
3/33 + 12
50 t
7140 _+ I I
16.5
15167 + 13.5
23*
Involved
T lymphocytes
13170 + 13
E A and E A C binding mononuclear ceils
1/35 27(+ 13) 8/40 :i: 12
*P < 0.05 t P < 0.01.
nuclear cells (ranging from 13 to 70%) showed surface bound Ig (B lymphocytes). In involved spleens, the mean percentage ofsplenic lymphocytes bearing Ig on the cell surface was 23%, with extreme values ranging from 7 to 40%. This difference is significant (P < 0.05). Table 2 also shows the percentages of non-T non-B mononuclear cells (nT nBmc). In noninvolved spleens the percentage was 31% (range: 0-63%) and in involved spleens 27% (range: 8-40%). No significant difference was detected. (B) Searchfor a correlation between immunological data and variables characterizing the disease This search led to three interesting observations: (1) We found a significantly higher percentage of splenic T lymphocytes in disseminated clinical stages III and IV than in cases of localized disease (stages I and II). We also found a slightly fewer mIg binding mononuclear cells, (B lymphocytes) EAC rosetteforming cells and nT n.Bmc in the generalized disease. However, these differences were not significant. No variation was detected in the percentage of splenic EA rosette-formlng cells in patients at different clinical stages of the disease. (2) We compared involved spleens in the localized and disseminated stages and found no difference in the percentages of T lymphocytes. Nor was any variation detected when comparing T cells of the non-involved spleens in each clinical group. (3) Finally, we found no clear-cut relationship in the spleen between the percentages of
the different mononuclear cell populations and the histological type of the tissue. DISCUSSION The results presented in this paper confirm our previous findings [17-21] as well as those of others [22, 23] that in Hodgkin's disease there is a significantly higher percentage of T lymphocytes present in spleens containing RSC, than in non-involved spleens. The mechanism of this increase is at present unclear. There are at least two possible explanations: (1) Since it has been claimed that RSC may be T cell derived, this increase in T mononuclear cells may correspond to an increase in RSC precursors. This hypothesis does not seem plausible, however, because recent reports, including our own [18-21], have suggested that RSC may in fact, be derived from B cells [24, 25] or from histiocytes [26, 27]. (2) This increase in T cells may correspond to a local immune reaction against RSC. This would explain the strikingly close contact seen with conventional [28] and scanning electron microscopy [18--20] between presumed T lymphocytes and RSC. These morphological findings do not, however, prove the existence of a specific immunological reaction. Our data regarding B lymphocytes confirm reports from other authors [29, 30] who found in using different methods a significant drop in the number of B cells in involved spleens. However, from our data we are unable to
358
A. Santoro, B. Caillou and D. Belpomme
deduce that there is a decrease in the absolute number of B lymphocytes in involved spleens in Hodgkin's disease. The absence of any significantdifference in the percentages of E A and E A C rosette-forming cells suggests that there is no difference in the percentage of cells related to the histiocytic ScI~eS. We showed a higher mean percentage of splenic E rosettes (T lymphocytcs) in disscminated clinical stages Ill and IV than in localized stages I and II. According to our findings,there isno differencein the percentage of E rosette forming cells between involved spleens, whether they arc localized or disscminatcd diseases. Our data showing a high mean percentage of splenic T lymphocytcs in the generalized clinical stages may in fact bc
explained by a larger number of involved spleens in the disseminated cases. There is however, no clear correlation between the increase of T lymphocytes in the spleen and the degree of dissemination of the disease. Our findings suggest that there is no correlation between the percentage of the different mononuclear cells and the histological type in the spleen; however, as our series was exclusively composed of NS and MC varieties, no definite conclusions can be drawn from these data. Finally, from this study it is suggested that the detection of an increased number of T lymphocytes in involved spleens may be clinically helpful in the evaluation of the clinical stage of Hodgkin's disease when pathological analysis remains difficult to interpret.
REFERENCES
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