Immunology 1976 31 803
Lymphocyte nucleolar activation as a marker of autoimmune disorders Il. OBSERVATIONS IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND OTHER CONNECTIVE TISSUE DISEASES
LIDKA KOR(AKOVA,* J. PEKAREK,t J. ROVENSKY,+ K. TRNAVSKY4, J. LUKACT & VPRA HA9KOVA * Transplantation Research Centre, Institute for Clinical and Experimental Medicine Prague, t Institute of Sera and Vaccines, Prague and t Research Institute of Rheumatic Diseases, Pijstacny, Czechoslovakia
Received 22 March 1976; accepted for publication 6 May 1976
Summary. An increased number of lymphocytes with active nucleoli was found in the peripheral blood of most patients suffering from connective tissue diseases associated with autoimmunity.
juvenile form and other connective tissue diseases with or without concomitant immune disorders, were investigated (Table 1). Illnesses were diagnosed in the Research Institute of Rheumatic Diseases according to internationally accepted criteria.
INTRODUCTION
Nucleolar test Dry unfixed-blood smears were stained with buffered toluidine blue, pH 5 (Smetana, Lejnar & Potmesil, 1969). One hundred lymphocytes were evaluated according to the morphology of their nucleoli. The cells with compact nucleoli and nucleoli with nucleolonemas, were considered active. Absolute numbers of active lymphocytes in 1 mm3 of blood were calculated from the values of the nucleolar test and from the total number of lymphocytes. Statistical evaluation of the results was performed by the Computer Centre of the Institute for Clinical and Experimental Medicine.
An increased number of active lymphocytes with nucleoli synthesizing RNA were found in blood and lymph nodes of aged NZB mice who developed an auto-immune disease (Korcakova', RovenskY, Pekairek & Haskova, 1976). This finding led us to follow activation of blood lymphocytes in humans suffering from various connective tissue diseases, especially those connected with auto-immune disorders. MATERIALS AND METHODS Patients Ninety patients suffering from systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), its Correspondence: Dr Jan Pekarek, Department of Experimental Immunology, Institute of Sera and Vaccines, Prague, Czechoslovakia.
RESULTS It can be seen from the summary of the results illustrated by Table 1 that in the majority of cases suffering from SLE, RA, RA juvenilis and scleroderma, the numbers of active lymphocytes were
803
804
Lidka Korc&kova et al. Table 1. Survey of values of active lymphocytes in humans with various connective tissue diseases
No. of cases
Group 1 2 3 4 5 6 7 8
Diagnosis SLE RA juvenilis RA Sclerodermia diffusa Psoriatic arthritis Other diseases with immune disorders* Coxarthrosis Controlst
positive/total
Active lymphocytes mean+ s.d. (percentage absolute number)
13/15 14/17 20/26
8-3+4-0 6-0+ 2-1 5-8+ 2-8
200-3+ 111-2 116-9+ 50 3 98-2+ 47*5
5/9
5-3+ 3-6
155-3+ 138-9
4/12
3-4+ 1-8
61P8+ 2-2
8/11 0/10
6-6+ 2-4 2-0+ 1-4 2-1+ 1-4
114-4+ 81[2 40-5+28-2 60-3+ 37-2
3/37
* Arteritis, polymyositis, dermatomyositis.
t Vertebrogenic syndrome, scoliosis, state after bone fracture, healthy children.
clearly elevated. The maximal increase was noted in patients with SLE where highly significant results were obtained. In ten patients with coxarthrosis and thirty-seven control patients suffering from connective tissue disorders having no immunological basis, the numbers of active lymphocytes fluctuated around normal values (Table 1, groups 7 and 8).
disorders are known to play a role (SLE, RA, RA juvenilis, scleroderma, etc.). The correlation between the values of active lymphocytes and the severity of the disease was not followed in the present study. However, it was previously found in one person that successful immunosuppressive therapy turned the positive value (12 per cent) into the negative one (1 per cent). It seems therefore useful to perform such comparative studies in the future.
DISCUSSION An increased number of active lymphocytes (i.e. lymphocytes with nucleoli synthesizing RNA) may reflect the activity of the immunological process, especially of the delayed hypersensitivity type (Pekarek, Veptekova' & Doutlik, 1972; Korcakova', 1973; Nekvasil & Pekairek, 1974). In humans the nucleolar activation was noted in patients with some infectious diseases (gramkova & Fortova, 1972), with cancer without metastases (Smetana Vlastiborova, Matejkova, Hondlik, Lejnar & Likovsky, 1976) and with kidney grafts at the time of rejection crises (Korcakova, Svobodova, Reneltova & Haskova', 1976). In the present paper activation of peripheral blood lymphocytes was followed in patients suffering from connective tissue diseases. Increased numbers of active lymphocytes were found in a majority of patients with diseases where autoimmune
REFERENCES KOR6AKOVA L. (1973) Blood lymphocyte nucleoli activation and skin allograft rejection in mice. Folia biol. (Praha), 19, 25. KOR6AKOVA L., ROVENSKY' J., PEKkREK J. & HASKOVA V. (1976) Lymphocyte nucleoli activation as a marker of autoimmune disorder development. I. Observations in NZB mice. Immunology, 30, 475. KOR6(KOvA L., SVOBODOVA J., RENELTOVA I. & HA§KOVJ V. (1976) Nucleolar test in a follow-up of renal allograft recipients. Correlation with the response to PHA. Transplantation, (In press.) NEKVASIL M. & PEKAREK J. (1974) Activation of the peripheral lymphocytes in the course of the immune response to transplanted SV40-transformed tumour cells in hamsters. Immunology, 27, 159. PEKAREK J., VEPAEKOvA A. & DOUTLIK S. (1972) Activation of peripheral lymphocytes in the course of experimental allergic encephalomyelitis in the guinea pig. Z. Immun. forsch. 143, 139.
Lymphocyte nucleoli activation SMETANA K., LEJNAR J. & POTMBAIL M. (1969) A further contribution to the demonstration of RNA and nucleoli in blood cells in smear preparations. Folia haemat. 91, 4. SMETANA K., VLASTIBOROVA A., MATtJKOVA E., HONDLIK J., LEJNAR J. & LIKOVSKi Z. (1976) Nucleoli of lymphocytes
I
805
in the peripheral blood of patients with bronchogenic lung and gastrointestinal cancer. Neoplasma, 23, 183. SRAMKOVJk L. & FORTOVA D. (1972) Investigation of lymphocytes activation in peripheral blood in infectious diseases. Z. Immun.-forsch. 144,129.
Lymphocyte nucleolar activation as a marker of autoimmune disorders Il. OBSERVATIONS IN PATIENTS WITH SYSTEMIC LUPUS ERYTHEMATOSUS AND OTHER CONNECTIVE TISSUE DISEASES
LIDKA KOR(AKOVA,* J. PEKAREK,t J. ROVENSKY,+ K. TRNAVSKY4, J. LUKACT & VPRA HA9KOVA * Transplantation Research Centre, Institute for Clinical and Experimental Medicine Prague, t Institute of Sera and Vaccines, Prague and t Research Institute of Rheumatic Diseases, Pijstacny, Czechoslovakia
Received 22 March 1976; accepted for publication 6 May 1976
Summary. An increased number of lymphocytes with active nucleoli was found in the peripheral blood of most patients suffering from connective tissue diseases associated with autoimmunity.
juvenile form and other connective tissue diseases with or without concomitant immune disorders, were investigated (Table 1). Illnesses were diagnosed in the Research Institute of Rheumatic Diseases according to internationally accepted criteria.
INTRODUCTION
Nucleolar test Dry unfixed-blood smears were stained with buffered toluidine blue, pH 5 (Smetana, Lejnar & Potmesil, 1969). One hundred lymphocytes were evaluated according to the morphology of their nucleoli. The cells with compact nucleoli and nucleoli with nucleolonemas, were considered active. Absolute numbers of active lymphocytes in 1 mm3 of blood were calculated from the values of the nucleolar test and from the total number of lymphocytes. Statistical evaluation of the results was performed by the Computer Centre of the Institute for Clinical and Experimental Medicine.
An increased number of active lymphocytes with nucleoli synthesizing RNA were found in blood and lymph nodes of aged NZB mice who developed an auto-immune disease (Korcakova', RovenskY, Pekairek & Haskova, 1976). This finding led us to follow activation of blood lymphocytes in humans suffering from various connective tissue diseases, especially those connected with auto-immune disorders. MATERIALS AND METHODS Patients Ninety patients suffering from systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), its Correspondence: Dr Jan Pekarek, Department of Experimental Immunology, Institute of Sera and Vaccines, Prague, Czechoslovakia.
RESULTS It can be seen from the summary of the results illustrated by Table 1 that in the majority of cases suffering from SLE, RA, RA juvenilis and scleroderma, the numbers of active lymphocytes were
803
804
Lidka Korc&kova et al. Table 1. Survey of values of active lymphocytes in humans with various connective tissue diseases
No. of cases
Group 1 2 3 4 5 6 7 8
Diagnosis SLE RA juvenilis RA Sclerodermia diffusa Psoriatic arthritis Other diseases with immune disorders* Coxarthrosis Controlst
positive/total
Active lymphocytes mean+ s.d. (percentage absolute number)
13/15 14/17 20/26
8-3+4-0 6-0+ 2-1 5-8+ 2-8
200-3+ 111-2 116-9+ 50 3 98-2+ 47*5
5/9
5-3+ 3-6
155-3+ 138-9
4/12
3-4+ 1-8
61P8+ 2-2
8/11 0/10
6-6+ 2-4 2-0+ 1-4 2-1+ 1-4
114-4+ 81[2 40-5+28-2 60-3+ 37-2
3/37
* Arteritis, polymyositis, dermatomyositis.
t Vertebrogenic syndrome, scoliosis, state after bone fracture, healthy children.
clearly elevated. The maximal increase was noted in patients with SLE where highly significant results were obtained. In ten patients with coxarthrosis and thirty-seven control patients suffering from connective tissue disorders having no immunological basis, the numbers of active lymphocytes fluctuated around normal values (Table 1, groups 7 and 8).
disorders are known to play a role (SLE, RA, RA juvenilis, scleroderma, etc.). The correlation between the values of active lymphocytes and the severity of the disease was not followed in the present study. However, it was previously found in one person that successful immunosuppressive therapy turned the positive value (12 per cent) into the negative one (1 per cent). It seems therefore useful to perform such comparative studies in the future.
DISCUSSION An increased number of active lymphocytes (i.e. lymphocytes with nucleoli synthesizing RNA) may reflect the activity of the immunological process, especially of the delayed hypersensitivity type (Pekarek, Veptekova' & Doutlik, 1972; Korcakova', 1973; Nekvasil & Pekairek, 1974). In humans the nucleolar activation was noted in patients with some infectious diseases (gramkova & Fortova, 1972), with cancer without metastases (Smetana Vlastiborova, Matejkova, Hondlik, Lejnar & Likovsky, 1976) and with kidney grafts at the time of rejection crises (Korcakova, Svobodova, Reneltova & Haskova', 1976). In the present paper activation of peripheral blood lymphocytes was followed in patients suffering from connective tissue diseases. Increased numbers of active lymphocytes were found in a majority of patients with diseases where autoimmune
REFERENCES KOR6AKOVA L. (1973) Blood lymphocyte nucleoli activation and skin allograft rejection in mice. Folia biol. (Praha), 19, 25. KOR6AKOVA L., ROVENSKY' J., PEKkREK J. & HASKOVA V. (1976) Lymphocyte nucleoli activation as a marker of autoimmune disorder development. I. Observations in NZB mice. Immunology, 30, 475. KOR6(KOvA L., SVOBODOVA J., RENELTOVA I. & HA§KOVJ V. (1976) Nucleolar test in a follow-up of renal allograft recipients. Correlation with the response to PHA. Transplantation, (In press.) NEKVASIL M. & PEKAREK J. (1974) Activation of the peripheral lymphocytes in the course of the immune response to transplanted SV40-transformed tumour cells in hamsters. Immunology, 27, 159. PEKAREK J., VEPAEKOvA A. & DOUTLIK S. (1972) Activation of peripheral lymphocytes in the course of experimental allergic encephalomyelitis in the guinea pig. Z. Immun. forsch. 143, 139.
Lymphocyte nucleoli activation SMETANA K., LEJNAR J. & POTMBAIL M. (1969) A further contribution to the demonstration of RNA and nucleoli in blood cells in smear preparations. Folia haemat. 91, 4. SMETANA K., VLASTIBOROVA A., MATtJKOVA E., HONDLIK J., LEJNAR J. & LIKOVSKi Z. (1976) Nucleoli of lymphocytes
I
805
in the peripheral blood of patients with bronchogenic lung and gastrointestinal cancer. Neoplasma, 23, 183. SRAMKOVJk L. & FORTOVA D. (1972) Investigation of lymphocytes activation in peripheral blood in infectious diseases. Z. Immun.-forsch. 144,129.
