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Phase variation in the modulation of the human immune response
213
Immunology Today, voL 4, No. 8, 1983
a n t i g e n s a p p e a r i n g o n h a e m o p o i e t i c cells, since t h e y are in d a i l y c o n t a c t w i t h t h e m a n d t h u s i m m u n o s e l e c t i o n a g a i n s t h a e m o p o i e t i c t u m o u r s will b e w e a k . F u r t h e r m o r e , w h e r e m a l i g n a n t t r a n s f o r m a t i o n results in t h e e x p r e s s i o n o f o n c o f o e t a l a n t i g e n s in the a b s e n c e o f a n y translation products derived from externally acquired n u c l e i c a c i d o r c a r c i n o g e n s , a fully d i f f e r e n t i a t e d l y m p h o cyte m i g h t b e e x p e c t e d to h a v e e x p e r i e n c e d s u c h a n t i g e n s d u r i n g its o w n p r o g r e s s to m a t u r i t y a n d t h e r e b y to h a v e b e e n p r o g r a m m e d to b e t o l e r a n t .
References
1 Boyse, E. A. and Old, L. J. (1969) Annu. Rev. Genet.3, 269 2 Reinherz, E. L. and Schlossman, S. F. (1980) Cell 19, 821 3 Lindahl, P., Gresser, I., Leary, P. and Tovey, M. (1976) Proc. NatlAcad. Sci. USA 13, 1284 4 Heron, I., Hoklan, M. and Berg, K. (1978) Proc. NatlAcad. Sci. USA 15, 6215 5 Berrone, O. R. and Milstein, C. (1982) EMBOJ. 1,345 6 Wallach, D., Fellous, M. and Revel, M. (1982) Nature(London) 299, 833 7 Atallah, A. M. and Strong, D. M. (1979) Int. Arch. AllergyAppl. Immunol. 60, 101 8 Duke, O., Panayi, G. S., Janossy, G. and Poulter, L. W. (1982) Clin. Exp. ImmunoL 49, 22
C,...-
)
Phase variation in the modulation of the h u m a n i m m u n e response SIR, M a r t i n Davies (Immunol. Today 1983, 4, 103) shows the importance o f documenting the phase variations in the modulation of i m m u n e responses, i.e. the same immunomodulating agent can induce both stimulation and depression of i m m u n e reactivity to antigenic stimuli, depending on small changes in the doses administered. As this phenomenon was documented only by the results of animal experiments, we should like to illustrate here an example of such variation in h u m a n beings. While attempting to correct the functional T-ceU deficiency associated with ageing 1, we treated 24 volunteers over 70 years with the synthetic pentapeptide TP-5 ( ' I m u n o x 'R) that in biological activity resembles the thymic h o r m o n e thymopoietin 2. TP-5 is non-toxic and rapidly degenerates into amino acids 3 (half-life: about 0.5 min). W e describe here the antibody responses to 500 fig keyhole limpet hemocyanin ( K L H ) , given subcutaneously. T h e 24 immunized subjects were assigned to one of four treatment groups in double-blind random fashion. In two of these parallel groups the patients underwent double-blind treatment with short (1 rain) intravenous injections of
9 Lampert, I. A., Switters, A. J. and Chisholm, P. M. (1981) Nature (London) 293, 149 10 Doherty, P. C., Blanden, R. V. and Zinkernagel, R. M. (1976) Transplant. Rev. 29, 89 11 Cikes, M. and Klein, G. (1972).]. Natl Cancer Inst. 49 1599 12 DiPersio, L. P., Weiss, M. A., Michael, J. G. and Pesce, A. J. (1982) Am. J Clin. PathoL 11, 100 13 Celis, E., Hale, A. H., Russel, J. H. and Eisen, H. N. (1979)J. ImmunoL 122,954 14 Trucco, M. M., Stocker,J. W. and Cepellini, R. (1978)Nature (London) 273, 666 15 Beverley, P. C. L. (1980) Transplantation and Clin~al Irnmunolog~ (Touraine, J. L., Traeger, J., B~tuel, H., Brochier, J., Dubernard, J. M., Revillard, J. P. and Trian, R., eds), Vol. XI, p. 87, Excerpta Medica, Amsterdam 16 Arce-Gomez, B., Jones, E. A., Barnstaple, C. J. et aL (1978) Tissue Antigens 11, 96 17 Solheim, B. G. and Thorsby, E. (1974) Tissue Antigens 4, 83 18 Brodsky, F. M., Parham, P., Barnstaple, C. J. et al. (1979)Immunol. Rev. 47, 3 19 Tjerlund, U. M. and Forsum, U. (1977)Acta Derm.-Venereol. 51,503 20 Sanderson, A. R. and Ward, P. J. (1981) Immunolog~ 44, 169 21 M611er,G. and M611er, E. (1975)J. Natl Cancer Inst. 55, 755 22 Basham, T. Y., Bourgeade, M. F., Creasey, A. A. and Merigan, T. C. (1982) Pr~'. NatlAcad. Sci. USA 19, 3265 23 Ruiter, R. J., Bhan, A. K., Harrist, T. J. et al. (1982)J. ImmungL 129, 28O8 24 Rosa, F., Berissa, H., Weissenbach, J. et al. (1983)EMBOJ. 2, 239 25 Holden, C. A., Shaw, M., McKee, P., Sanderson, A. R. and MacDonald, D. M. J. Am. Acad. Damatol. (in press)
either 50 m g TP-5 (IVS) or placebo (P). T h e patients in the other two groups received prolonged (10 min) intravenous (IVL) and subcutaneous (SC) injections, respectively, of the same TP-5 doses. T h e injections were regularly repeated six times within 14 days; blood samples were taken at weekly intervals for 3 weeks. T h e influence of the adjuvant
treatment (TP-5 or placebo) on the antibody responses was measured by radioimmunoassay 4 modified for K L H specific I g M and IgG antibodies. T h e titers were expressed by reference to a standard curve obtained by dilutions o f a h y p e r i m m u n e serum. T h e level of the specific antibodies increased during the study period, but
TITER
CIk~ IgM
P
SC
IVL
IVS
Fig. 1. A n t i - K L H antibodies at the third w e e k after vaccination.
Adjuvant treatment: P, placebo; SC, 50 mg TP-5 subcutaneously; IVL, .50 mg TP-5 intravenously, applied over 10 min; IVS, 50 mg TP-5 intravenously (bolus). © ElsevierBiomedicaJPress19830167-491918310000-0000151.00
Immunology Today, vol. 4, No. 8, 1983
214 T A B L E I. Intergroup differences in the anti-KLH IgG and IgM titers. Antibody response comparing the treatment groups
Week 1
Week 2
Week 3
S.c. TP-5 > placebo (SO > P) S.c. TP-5 > i.v. TP-5 (SC > IVS)
Not significant
IgG (P < 0.1 )
IgG (P < 0.05)
IgG (P< 0.05) IgM (P < 0.1)
IgG (P < 0.05) IgM (P < 0.05)
S.c. TP-5 > i.v. long TP-5 (SC > IVL)
Not significant
Not significant
IgG (P < 0.05) IgM (not significant) IgG (P< 0.1) IgM ( P < 0.05)
Placebo > i.v. TP-5 (P > IVS)
IgG (P < 0.05)
Not significant
Not significant
some differences were observed among the treated groups. The responses in week 3 are presented in Fig. 1. The SC group reached the highest means of specific IgM and IgG anti-KLH antibody levels. These levels were significantly different (P < 0.05, by one-tailed Mann-Whitney test) from those of the P group and, moreover, the IVS group for IgG and the IVL group for IgM, respectively. The statistically significant intergroup differences observed during the study in the anti-KLH IgG and IgM (Table I) indicate the phase-modulating effect of TP-5. While the specific immune response in the IV groups was almost
always lower (significantly lower IgG levels in week 1 in the IVS group; P < 0.05) than in the placebo group, the SC group invariably showed higher production of both these specific antibodies (significantly higher IgG levels after weeks 2 and 3; P < 0.1 and < 0.05, respectively). Moreover, the SC group at each rating showed for at least one parameter a significantly higher response ( P < 0.05) than the IV groups. These different patterns of response associated with the different routes of application of TP-5 can be related to differences in drug availability with respect to site of action, concentration, and duration of contact with appropriate
target cells. Intravenously injected doses proved to be biologically more active than the same doses administered subcutaneously (T. Audhya and G. Goldstein, unpublished observations). Therefore, we feel that TP-5 may modulate immune reactivity, depending upon the route of administration or the dose level. This investigation not only demonstrates the phenomenon of phase variation by a thymic hormone preparation, but also gives additional evidence of the immunomodulating character of TP-5. J. DUCHATEAU Department of Immunology, H@ital Universitaire, Saint-Pierre (ULB), Brussels, Belgium. G. DELESPESSE Department of Immunology, University of Manitoba, Winnipeg, Canada. K. BOLLA Clinical R&D, Technical Center, CILAG Ltd, Schaffhausen, Switzerland.
References 1 Makinodan, T. and Kay, M. B. (1980)Adv. Immunol. 29, 287-330 2 Goldstein,G., Scheid, M. P., Boyse, E. A. et a/. (1979) ScLm¢~(Valparaiso) 204, 1309 3 Tischio,J. P., Patrick, J. E., Weintraub, H. S. et aL (1979)Int. J. Pept. ProteinRes. 14, 479 4 Duchateau, J., Delepesse, G., Vryens, R. and Collet, H. (1981) Am. J. Med. 70, 1001-1004
Immunology Today, voL 4, No. 8, 1983
a n t i g e n s a p p e a r i n g o n h a e m o p o i e t i c cells, since t h e y are in d a i l y c o n t a c t w i t h t h e m a n d t h u s i m m u n o s e l e c t i o n a g a i n s t h a e m o p o i e t i c t u m o u r s will b e w e a k . F u r t h e r m o r e , w h e r e m a l i g n a n t t r a n s f o r m a t i o n results in t h e e x p r e s s i o n o f o n c o f o e t a l a n t i g e n s in the a b s e n c e o f a n y translation products derived from externally acquired n u c l e i c a c i d o r c a r c i n o g e n s , a fully d i f f e r e n t i a t e d l y m p h o cyte m i g h t b e e x p e c t e d to h a v e e x p e r i e n c e d s u c h a n t i g e n s d u r i n g its o w n p r o g r e s s to m a t u r i t y a n d t h e r e b y to h a v e b e e n p r o g r a m m e d to b e t o l e r a n t .
References
1 Boyse, E. A. and Old, L. J. (1969) Annu. Rev. Genet.3, 269 2 Reinherz, E. L. and Schlossman, S. F. (1980) Cell 19, 821 3 Lindahl, P., Gresser, I., Leary, P. and Tovey, M. (1976) Proc. NatlAcad. Sci. USA 13, 1284 4 Heron, I., Hoklan, M. and Berg, K. (1978) Proc. NatlAcad. Sci. USA 15, 6215 5 Berrone, O. R. and Milstein, C. (1982) EMBOJ. 1,345 6 Wallach, D., Fellous, M. and Revel, M. (1982) Nature(London) 299, 833 7 Atallah, A. M. and Strong, D. M. (1979) Int. Arch. AllergyAppl. Immunol. 60, 101 8 Duke, O., Panayi, G. S., Janossy, G. and Poulter, L. W. (1982) Clin. Exp. ImmunoL 49, 22
C,...-
)
Phase variation in the modulation of the h u m a n i m m u n e response SIR, M a r t i n Davies (Immunol. Today 1983, 4, 103) shows the importance o f documenting the phase variations in the modulation of i m m u n e responses, i.e. the same immunomodulating agent can induce both stimulation and depression of i m m u n e reactivity to antigenic stimuli, depending on small changes in the doses administered. As this phenomenon was documented only by the results of animal experiments, we should like to illustrate here an example of such variation in h u m a n beings. While attempting to correct the functional T-ceU deficiency associated with ageing 1, we treated 24 volunteers over 70 years with the synthetic pentapeptide TP-5 ( ' I m u n o x 'R) that in biological activity resembles the thymic h o r m o n e thymopoietin 2. TP-5 is non-toxic and rapidly degenerates into amino acids 3 (half-life: about 0.5 min). W e describe here the antibody responses to 500 fig keyhole limpet hemocyanin ( K L H ) , given subcutaneously. T h e 24 immunized subjects were assigned to one of four treatment groups in double-blind random fashion. In two of these parallel groups the patients underwent double-blind treatment with short (1 rain) intravenous injections of
9 Lampert, I. A., Switters, A. J. and Chisholm, P. M. (1981) Nature (London) 293, 149 10 Doherty, P. C., Blanden, R. V. and Zinkernagel, R. M. (1976) Transplant. Rev. 29, 89 11 Cikes, M. and Klein, G. (1972).]. Natl Cancer Inst. 49 1599 12 DiPersio, L. P., Weiss, M. A., Michael, J. G. and Pesce, A. J. (1982) Am. J Clin. PathoL 11, 100 13 Celis, E., Hale, A. H., Russel, J. H. and Eisen, H. N. (1979)J. ImmunoL 122,954 14 Trucco, M. M., Stocker,J. W. and Cepellini, R. (1978)Nature (London) 273, 666 15 Beverley, P. C. L. (1980) Transplantation and Clin~al Irnmunolog~ (Touraine, J. L., Traeger, J., B~tuel, H., Brochier, J., Dubernard, J. M., Revillard, J. P. and Trian, R., eds), Vol. XI, p. 87, Excerpta Medica, Amsterdam 16 Arce-Gomez, B., Jones, E. A., Barnstaple, C. J. et aL (1978) Tissue Antigens 11, 96 17 Solheim, B. G. and Thorsby, E. (1974) Tissue Antigens 4, 83 18 Brodsky, F. M., Parham, P., Barnstaple, C. J. et al. (1979)Immunol. Rev. 47, 3 19 Tjerlund, U. M. and Forsum, U. (1977)Acta Derm.-Venereol. 51,503 20 Sanderson, A. R. and Ward, P. J. (1981) Immunolog~ 44, 169 21 M611er,G. and M611er, E. (1975)J. Natl Cancer Inst. 55, 755 22 Basham, T. Y., Bourgeade, M. F., Creasey, A. A. and Merigan, T. C. (1982) Pr~'. NatlAcad. Sci. USA 19, 3265 23 Ruiter, R. J., Bhan, A. K., Harrist, T. J. et al. (1982)J. ImmungL 129, 28O8 24 Rosa, F., Berissa, H., Weissenbach, J. et al. (1983)EMBOJ. 2, 239 25 Holden, C. A., Shaw, M., McKee, P., Sanderson, A. R. and MacDonald, D. M. J. Am. Acad. Damatol. (in press)
either 50 m g TP-5 (IVS) or placebo (P). T h e patients in the other two groups received prolonged (10 min) intravenous (IVL) and subcutaneous (SC) injections, respectively, of the same TP-5 doses. T h e injections were regularly repeated six times within 14 days; blood samples were taken at weekly intervals for 3 weeks. T h e influence of the adjuvant
treatment (TP-5 or placebo) on the antibody responses was measured by radioimmunoassay 4 modified for K L H specific I g M and IgG antibodies. T h e titers were expressed by reference to a standard curve obtained by dilutions o f a h y p e r i m m u n e serum. T h e level of the specific antibodies increased during the study period, but
TITER
CIk~ IgM
P
SC
IVL
IVS
Fig. 1. A n t i - K L H antibodies at the third w e e k after vaccination.
Adjuvant treatment: P, placebo; SC, 50 mg TP-5 subcutaneously; IVL, .50 mg TP-5 intravenously, applied over 10 min; IVS, 50 mg TP-5 intravenously (bolus). © ElsevierBiomedicaJPress19830167-491918310000-0000151.00
Immunology Today, vol. 4, No. 8, 1983
214 T A B L E I. Intergroup differences in the anti-KLH IgG and IgM titers. Antibody response comparing the treatment groups
Week 1
Week 2
Week 3
S.c. TP-5 > placebo (SO > P) S.c. TP-5 > i.v. TP-5 (SC > IVS)
Not significant
IgG (P < 0.1 )
IgG (P < 0.05)
IgG (P< 0.05) IgM (P < 0.1)
IgG (P < 0.05) IgM (P < 0.05)
S.c. TP-5 > i.v. long TP-5 (SC > IVL)
Not significant
Not significant
IgG (P < 0.05) IgM (not significant) IgG (P< 0.1) IgM ( P < 0.05)
Placebo > i.v. TP-5 (P > IVS)
IgG (P < 0.05)
Not significant
Not significant
some differences were observed among the treated groups. The responses in week 3 are presented in Fig. 1. The SC group reached the highest means of specific IgM and IgG anti-KLH antibody levels. These levels were significantly different (P < 0.05, by one-tailed Mann-Whitney test) from those of the P group and, moreover, the IVS group for IgG and the IVL group for IgM, respectively. The statistically significant intergroup differences observed during the study in the anti-KLH IgG and IgM (Table I) indicate the phase-modulating effect of TP-5. While the specific immune response in the IV groups was almost
always lower (significantly lower IgG levels in week 1 in the IVS group; P < 0.05) than in the placebo group, the SC group invariably showed higher production of both these specific antibodies (significantly higher IgG levels after weeks 2 and 3; P < 0.1 and < 0.05, respectively). Moreover, the SC group at each rating showed for at least one parameter a significantly higher response ( P < 0.05) than the IV groups. These different patterns of response associated with the different routes of application of TP-5 can be related to differences in drug availability with respect to site of action, concentration, and duration of contact with appropriate
target cells. Intravenously injected doses proved to be biologically more active than the same doses administered subcutaneously (T. Audhya and G. Goldstein, unpublished observations). Therefore, we feel that TP-5 may modulate immune reactivity, depending upon the route of administration or the dose level. This investigation not only demonstrates the phenomenon of phase variation by a thymic hormone preparation, but also gives additional evidence of the immunomodulating character of TP-5. J. DUCHATEAU Department of Immunology, H@ital Universitaire, Saint-Pierre (ULB), Brussels, Belgium. G. DELESPESSE Department of Immunology, University of Manitoba, Winnipeg, Canada. K. BOLLA Clinical R&D, Technical Center, CILAG Ltd, Schaffhausen, Switzerland.
References 1 Makinodan, T. and Kay, M. B. (1980)Adv. Immunol. 29, 287-330 2 Goldstein,G., Scheid, M. P., Boyse, E. A. et a/. (1979) ScLm¢~(Valparaiso) 204, 1309 3 Tischio,J. P., Patrick, J. E., Weintraub, H. S. et aL (1979)Int. J. Pept. ProteinRes. 14, 479 4 Duchateau, J., Delepesse, G., Vryens, R. and Collet, H. (1981) Am. J. Med. 70, 1001-1004