Asia-Oceania J . Obstet. Gynaecol. Val. 18, No. 2; 171-175 1992
Cell-Mediated Immunity in Cervical Intraepithelial Neoplasia
Tchan K. Park, So0 N. Kim, Dong H. Choi, and Chang H. Lee Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul, Korea
Abstract Various immune parameters, i.e., peripheral T and B cell distribution, T cell subpopulation, interleukin 2 (IL-2) activity and in vitro lymphocyte responses to phytohemagglutinin (PHA) and concanavalin A (Con A), were investigated in 24 pretreated patients with cervical intraepithelial neoplasia (CIN) and in 35 normal controls. Of the measured parameters, lymphocyte response to PHA was significantly lower in the CIN than control group (P0.05 interleukin-2 Data are expressed as meanfSD.
a)
Results The difference in relation to the mean number and percentage of T and B cells, respectively, in the peripheral blood of the CIN and control groups was not significant (P>O.OS) (Table 1). No significant difference in any of the percentage of CD4+ or CD8+ T cells, and T cell helper-to-suppressor subset ratio (CD4+/CD8+) was present between the CIN and control group (P>O.OS) (Table 2). Lymphocyte response to PHA was significantly lower in the CIN than the control group (7.2-113.2 vs. 20.9-cl4.8) (PO.O5) (Table 3). The mean IL-2 production was similar in both groups (P>O.OS) (Table 4).
Discussion This study shares some features with our previous study161 done on CMI in invasive cervical carcinoma (ICC) in that the same laboratory methodology and the same data set for the controls were used. In this study, the CIN group replaced ICC group as a new study group, and comparison with the control group was made in respect to the measured immune parameters. Unlike the previous studyle)where the values of nearly all parameters of CMI
measured were significantly lower in the ICC group than the controls, the blastogenic response of lymphocyte to PHA in the CIN group among the parameters measured was the only parameter associated with lower value of statistical significance than the controls in the current study. Although T cell population was not significantly different in both the CIN and control groups, there are studies1J7) describing significantly decreased overall T cell population in CIN patients compared with the controls. It therefore seems that whether or not T cell decrease actually accompanies CIN patients needs further investigation involving a larger sample size. One interesting facet of our current results is that, though not significantly fewer than those of the controls, T cell population and percentage of one of its subset, CD4+, were nonetheless all smaller in the CIN patients as well as the ratio of CD4+ to CD8+. This raises the possibility of attaining statistical significance by reducing the potential “type 11” error by incorporating additional CIN cases into study. Such speculation is made stronger when we take a note of significantly decreased lymphocyte response to PHA in the CIN patients studied. It is perhaps the functional disturbance of T cells that precedes any visible change in its numerical decrease. PHA is known to preferentially stimulates helper T cells. Our finding of decreased PHA response in the absence of decrease in CD4+ again makes it plausible the possibility of functional disturbance of CD4+ prior to its numerical decline. It may be that CIN patients with more decreased PHA response show more rapid progression to invasive cancer and for that matter decreased PHA response may serve as an early criterion for more tight follow-up or early intervention. No significant difference of response to Con A in the CIN patients contrasts with our previous studyle) involving ICC patients which showed uniformly low Con A response of statistical significance regardless of cancer stage, as did several similar ~ t u d i e s . ~ * J ~Daunter J~) et aZ.sO)observed in their 21 ICC patients, a low response to PHA but a similar response to 173
T . K . PARK BT AL.
Con A compared with those of the controls. In this vein, Con A response may be considered a late immune parameter, the value of which drops significantly only with CMI depression that accompanies decreased T cell population, notably CD4+. Unchanged IL-2 activity probably has similar connotation given to Con A response in that it may not be a good indicator of subtle and early CMI change. With advancing CMI depression usually associated with cancer progression, as probably was the case in some studies including our previous one,l'3BZ1)IL-2 activity may drop significantly. CD4+ and CD8+ cells have been shown to produce IL2 in e ) i t ~ o , 2 2 9 2 S )and this possibly is a reason for our normal finding on IL-2 activity. We regret that CIN grades of our patients were not available to determine the grade-specific measurement of the studied parameters. Perhaps, a future study may address this issue.
References 1. Ishiguro T, Sugitachi I, Katoh K. T and B
2.
3.
4.
5.
6.
7.
174
lymphocytes in patients with squamous cell carcinoma of the uterine cervix. Gynecol Oncol 1980; 9: 80-85 Levy S, Kopersztych S, Musatti CC, Souen JS, Savatore CA, Mendes NF. Cellular immunity in squamous cell carcinoma of the uterine cervix. A m J Obstet Gynecol 1978; 130: 160-1 64 Bigbee WL, Jensen RH. Characterization of plasminogen activator in human cervical cells. Biochem Biophys Acta 1978; 540: 285-294 Chen SS, Koffler D, Cohen CJ. Cellular hypersensitivity in patients with squamous cell carcinoma of the cervix. A m J Obstet Gynecol 1975;121:91-95 Chiang WT, Wei PY, Alexander ER. Circulatory and cellular immune responses to squamous cell carcinoma of the uterine cervix. A m J Obstet Gynecoll976; 126: 116 Cocchiara R, Tarro G, Flaminio G, Gioia MD, Geraci D. Enzyme-linked immunosorbent assay for herpes simplex virus tumor-associated antigen. Cancer 1980; 45 : 938-942 DiSaia PJ, Sinkovics JG, Rutledge FN, Smith JP. Cell-mediated immunity to human malignant cells. A m J Obstet Gynecol 1972; 114: 979-989
8. Goldenberg DM, Garner TF, Pant KD, van Nagell JR. Jr. Identification of beta-oncofetal antigen in cervical squamous cancer and its demonstration in neoplastic and normal tissues. Cancer Res 1978; 38: 1246-1249 9. Hagen C, Froland A, Weberg E. Lymphocyte transformation in cancer patients. Lancet 1972; 1: 1340 10. Kato H, Morioka H, Aramaki S, Tamai K, Torigoe T. Prognostic significance of the tumor antigen TA-4 in squamous cell carcinoma of the uterine cervix. A m J Obstet Gynecol 1983; 145: 350-354 11. Kietlinska 2. T and B lymphocyte counts and blast transformation in patients with Stage I cervical cancer. Gynecol Oncol 1984; 18(2): 247-256 12. Mashiba H, Matsunaga K, Ueno M, Jimi S. Cell-mediated cytotoxicity in witro of human lymphocytes against a cervical cancer cell line. Gann 1977; 68: 53-58 13. Micksche M, Luger TH, Michalica W, Tatra G. Investigations on general immune reactivity in untreated cervical cancer patients. Oncology 1978; 35: 206-209 14. Robbins DS, Fudenberg HH. Human lymphocyte subpopulations in metastatic neoplasia: Six years later. N Engl J Med 1983; 308: 1595-1 597 15. Schantz SP, Poisson L, Romsdahl MM, Byers RM. Natural killer cell activity and head and neck cancer. Proc A m Assoc Can Res 1985; 26: 308 16. Park TK, Kim SN. Cell-mediated immunity in patients with invasive carcinoma of the cervix. Yonsei Med J 1987; 30: 164-172 17. Castello G, Esposito G, Stellato G, Dalla Mora L, Abate G, German0 A. Immunological abnormalities in patients with cervical carcinoma. Gynecol Oncoll986; 25(1): 61-64 18. Rand RJ, Jenkins DM, Bulmer R. T-and Blymphocyte subpopulations following radiotherapy for invasive squamous carcinoma of the cervix. Clin Exp Immunol 1977; 30: 159165 19. Rey A, Klein B, Rucheton M, Caraux J, Zagury D, Thierry C, Serrou B. Human autologoua rosettes IV. Their relation with interleukin 2 activity production and natural killer cells in cancer patients. Cellular Immunol 1984; 86: 155-164 20. Daunter B, Khoo SK, Mackay EV. Lymphocyte response to plant mitogens 11. The response of lymphocytes from women with carcinoma of the cervix to phytohemaggluthh-P,
CELL-MBDIATED IMMUNITY IN CIN
concanavalin A, and pokeweed. Gynecol Oncol 1979; 7: 314-317 21. Dawson DE, Everts EC, Vetto RM, Burger DR. Assessment of immunocompetent cells in patients with head and neck squamous cell carcinoma. Ann Otol Rhino1 Laryngol198.5; 94: 342-345 22. Andrus L, Granelli-Piperno A, Reich E. Cyto-
toxic T cells both produce and respond to interleukin 2. J Ex# Med 1984; 59: 647-652 23. Luger TA, Smolen JS, Chused TM, Steinberg AD, Oppenheim JJ. Human lymphocytes with either the OKT4 or OKT8 phenotype produce interleukin 2 in culture. J CZin Invest 1982; 70: 470473
175
Cell-Mediated Immunity in Cervical Intraepithelial Neoplasia
Tchan K. Park, So0 N. Kim, Dong H. Choi, and Chang H. Lee Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul, Korea
Abstract Various immune parameters, i.e., peripheral T and B cell distribution, T cell subpopulation, interleukin 2 (IL-2) activity and in vitro lymphocyte responses to phytohemagglutinin (PHA) and concanavalin A (Con A), were investigated in 24 pretreated patients with cervical intraepithelial neoplasia (CIN) and in 35 normal controls. Of the measured parameters, lymphocyte response to PHA was significantly lower in the CIN than control group (P0.05 interleukin-2 Data are expressed as meanfSD.
a)
Results The difference in relation to the mean number and percentage of T and B cells, respectively, in the peripheral blood of the CIN and control groups was not significant (P>O.OS) (Table 1). No significant difference in any of the percentage of CD4+ or CD8+ T cells, and T cell helper-to-suppressor subset ratio (CD4+/CD8+) was present between the CIN and control group (P>O.OS) (Table 2). Lymphocyte response to PHA was significantly lower in the CIN than the control group (7.2-113.2 vs. 20.9-cl4.8) (PO.O5) (Table 3). The mean IL-2 production was similar in both groups (P>O.OS) (Table 4).
Discussion This study shares some features with our previous study161 done on CMI in invasive cervical carcinoma (ICC) in that the same laboratory methodology and the same data set for the controls were used. In this study, the CIN group replaced ICC group as a new study group, and comparison with the control group was made in respect to the measured immune parameters. Unlike the previous studyle)where the values of nearly all parameters of CMI
measured were significantly lower in the ICC group than the controls, the blastogenic response of lymphocyte to PHA in the CIN group among the parameters measured was the only parameter associated with lower value of statistical significance than the controls in the current study. Although T cell population was not significantly different in both the CIN and control groups, there are studies1J7) describing significantly decreased overall T cell population in CIN patients compared with the controls. It therefore seems that whether or not T cell decrease actually accompanies CIN patients needs further investigation involving a larger sample size. One interesting facet of our current results is that, though not significantly fewer than those of the controls, T cell population and percentage of one of its subset, CD4+, were nonetheless all smaller in the CIN patients as well as the ratio of CD4+ to CD8+. This raises the possibility of attaining statistical significance by reducing the potential “type 11” error by incorporating additional CIN cases into study. Such speculation is made stronger when we take a note of significantly decreased lymphocyte response to PHA in the CIN patients studied. It is perhaps the functional disturbance of T cells that precedes any visible change in its numerical decrease. PHA is known to preferentially stimulates helper T cells. Our finding of decreased PHA response in the absence of decrease in CD4+ again makes it plausible the possibility of functional disturbance of CD4+ prior to its numerical decline. It may be that CIN patients with more decreased PHA response show more rapid progression to invasive cancer and for that matter decreased PHA response may serve as an early criterion for more tight follow-up or early intervention. No significant difference of response to Con A in the CIN patients contrasts with our previous studyle) involving ICC patients which showed uniformly low Con A response of statistical significance regardless of cancer stage, as did several similar ~ t u d i e s . ~ * J ~Daunter J~) et aZ.sO)observed in their 21 ICC patients, a low response to PHA but a similar response to 173
T . K . PARK BT AL.
Con A compared with those of the controls. In this vein, Con A response may be considered a late immune parameter, the value of which drops significantly only with CMI depression that accompanies decreased T cell population, notably CD4+. Unchanged IL-2 activity probably has similar connotation given to Con A response in that it may not be a good indicator of subtle and early CMI change. With advancing CMI depression usually associated with cancer progression, as probably was the case in some studies including our previous one,l'3BZ1)IL-2 activity may drop significantly. CD4+ and CD8+ cells have been shown to produce IL2 in e ) i t ~ o , 2 2 9 2 S )and this possibly is a reason for our normal finding on IL-2 activity. We regret that CIN grades of our patients were not available to determine the grade-specific measurement of the studied parameters. Perhaps, a future study may address this issue.
References 1. Ishiguro T, Sugitachi I, Katoh K. T and B
2.
3.
4.
5.
6.
7.
174
lymphocytes in patients with squamous cell carcinoma of the uterine cervix. Gynecol Oncol 1980; 9: 80-85 Levy S, Kopersztych S, Musatti CC, Souen JS, Savatore CA, Mendes NF. Cellular immunity in squamous cell carcinoma of the uterine cervix. A m J Obstet Gynecol 1978; 130: 160-1 64 Bigbee WL, Jensen RH. Characterization of plasminogen activator in human cervical cells. Biochem Biophys Acta 1978; 540: 285-294 Chen SS, Koffler D, Cohen CJ. Cellular hypersensitivity in patients with squamous cell carcinoma of the cervix. A m J Obstet Gynecol 1975;121:91-95 Chiang WT, Wei PY, Alexander ER. Circulatory and cellular immune responses to squamous cell carcinoma of the uterine cervix. A m J Obstet Gynecoll976; 126: 116 Cocchiara R, Tarro G, Flaminio G, Gioia MD, Geraci D. Enzyme-linked immunosorbent assay for herpes simplex virus tumor-associated antigen. Cancer 1980; 45 : 938-942 DiSaia PJ, Sinkovics JG, Rutledge FN, Smith JP. Cell-mediated immunity to human malignant cells. A m J Obstet Gynecol 1972; 114: 979-989
8. Goldenberg DM, Garner TF, Pant KD, van Nagell JR. Jr. Identification of beta-oncofetal antigen in cervical squamous cancer and its demonstration in neoplastic and normal tissues. Cancer Res 1978; 38: 1246-1249 9. Hagen C, Froland A, Weberg E. Lymphocyte transformation in cancer patients. Lancet 1972; 1: 1340 10. Kato H, Morioka H, Aramaki S, Tamai K, Torigoe T. Prognostic significance of the tumor antigen TA-4 in squamous cell carcinoma of the uterine cervix. A m J Obstet Gynecol 1983; 145: 350-354 11. Kietlinska 2. T and B lymphocyte counts and blast transformation in patients with Stage I cervical cancer. Gynecol Oncol 1984; 18(2): 247-256 12. Mashiba H, Matsunaga K, Ueno M, Jimi S. Cell-mediated cytotoxicity in witro of human lymphocytes against a cervical cancer cell line. Gann 1977; 68: 53-58 13. Micksche M, Luger TH, Michalica W, Tatra G. Investigations on general immune reactivity in untreated cervical cancer patients. Oncology 1978; 35: 206-209 14. Robbins DS, Fudenberg HH. Human lymphocyte subpopulations in metastatic neoplasia: Six years later. N Engl J Med 1983; 308: 1595-1 597 15. Schantz SP, Poisson L, Romsdahl MM, Byers RM. Natural killer cell activity and head and neck cancer. Proc A m Assoc Can Res 1985; 26: 308 16. Park TK, Kim SN. Cell-mediated immunity in patients with invasive carcinoma of the cervix. Yonsei Med J 1987; 30: 164-172 17. Castello G, Esposito G, Stellato G, Dalla Mora L, Abate G, German0 A. Immunological abnormalities in patients with cervical carcinoma. Gynecol Oncoll986; 25(1): 61-64 18. Rand RJ, Jenkins DM, Bulmer R. T-and Blymphocyte subpopulations following radiotherapy for invasive squamous carcinoma of the cervix. Clin Exp Immunol 1977; 30: 159165 19. Rey A, Klein B, Rucheton M, Caraux J, Zagury D, Thierry C, Serrou B. Human autologoua rosettes IV. Their relation with interleukin 2 activity production and natural killer cells in cancer patients. Cellular Immunol 1984; 86: 155-164 20. Daunter B, Khoo SK, Mackay EV. Lymphocyte response to plant mitogens 11. The response of lymphocytes from women with carcinoma of the cervix to phytohemaggluthh-P,
CELL-MBDIATED IMMUNITY IN CIN
concanavalin A, and pokeweed. Gynecol Oncol 1979; 7: 314-317 21. Dawson DE, Everts EC, Vetto RM, Burger DR. Assessment of immunocompetent cells in patients with head and neck squamous cell carcinoma. Ann Otol Rhino1 Laryngol198.5; 94: 342-345 22. Andrus L, Granelli-Piperno A, Reich E. Cyto-
toxic T cells both produce and respond to interleukin 2. J Ex# Med 1984; 59: 647-652 23. Luger TA, Smolen JS, Chused TM, Steinberg AD, Oppenheim JJ. Human lymphocytes with either the OKT4 or OKT8 phenotype produce interleukin 2 in culture. J CZin Invest 1982; 70: 470473
175
