Dermatológica 157: 28-32 (1978)
Immunological Studies in Psoriasis T.R. Beim, S. Sehgal, S. Sengupta , S. K aur and B.K. A ikat Departments of Dermatology, Immunopathology, and Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh
Key Words. Psoriasis • Immunoglobulins IgG, IgM, IgA, IgD • Antiglobulins Abstract. The mean values of serum immunoglobulins A, G, M and D levels in 42 psoriasis patients showed no significant difference from those of control subjects. Analysis of the same sera for the presence of antiglobulin antibodies also yielded negative results. Antinuclear factor could be demonstrated in only 4.8% of the cases. The possibility that serum antiglobulins get drained into the lesions making their detection in the serum difficult, is supported by our preliminary findings of focal deposition in the stratum corneum of immune complexes of IgG, IgM and complement in tissue sections of psoriatic lesions from these patients.
Received: September 19, 1977; accepted: November 4, 1977.
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
The autoimmune hypothesis regarding etiopathogenesis of psoriasis is largely based on the demonstration of serum antibodies to stratum corneum and complement-mediated antigen-antibody complexes in the superficial layers of the psoriatic epidermis [1, 6-8], The immune complexes, perhaps, initiate a chain of events eliciting chemotactic cellular reaction resulting in the histogenesis of psoriatic lesion. Antiglobulin factors have been shown in the psoriatic lesion [2, 5] and studies on serum immunoglobulins have yielded rather contradictory results. Variously, the serum IgG has been reported to be normal [17], low [3, 9] or high [4, 11]. Serum IgM is reported to be either normal [4, 11, 17] or low [3, 9], Apart from a solitary report of low serum IgA [9], it has been shown by many workers to be raised [3, 4, 11, 17], Results of serum IgD estimations in psoriasis are not available.
Immunological Studies in Psoriasis
29
Anti-IgG activity has been shown to be present on blood lymphocytes from patients with psoriasis [13]. The success to demonstrate antiglobulin factors in the serum, however, appears to be related to the specificity of anti globulin and the technique employed for its detection [2], We have attempted to evaluate serum immunoglobulins G, M, A and D by conventional methods and antiglobulin factors in the serum employing techniques, both for agglutinating and nonagglutinating antibodies. The results are presented herein.
Materials and Methods The investigation was carried out on 42 patients (12 female and 30 male) with psoriasis; the age ranging from 15 to 55 years and the duration of the disease varying from 2 months to 30 years. The body surface area affected varied from 20 to 100%; arthralgia was present in 6 patients including 1 having deforming arthropathy of the hands and feet. 37 patients had received topical tar applications; 6 of them also received topical corticosteroid treatment at some time during their illness and the other 5 had no treatment prior to the study. None had received internal corticosteroids or cytotoxic drugs. The control group comprised 20 age-matched normal healthy individuals. Serum Immunoglobulin Estimation The quantitative determination of serum IgG, IgM, IgA and IgD were carried out by the radial immunodiffusion technique [10] using monospecific antisera obtained from Hyland Laboratory, USA. Reference standard serum for immunoglobulins was obtained from WHO (courtesy D.S. R owe). Antinuclear Antibodies The conventional indirect IF technique was employed. Briefly 4 ¡xm thick air-dried cryostat sections of rat liver were layered with 1:10 dilution patients’ serum for 30 min at 37 °C, washed in several changes of phosphate-buffered saline (pH 7.4) and treated with fluorescein-labelled polyvalent antihuman immunoglobulin antiserum (Burroughs-Wellcome) at a dilution of 1 in 10 for another 30 min at 37°C. The sections were washed in phosphate-buffered saline and mounted in buffered glycerin and examined under fluores cent light.
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
Antiglobulin Factor Antiglobulin factors were searched for in the patients sera employing 2 different tech niques: (A) Tanned red cell (TRC) hemagglutination method employing denatured human y-globulin (Curewell India Ltd.). Sheep red cells collected in Alsevers solution, washed in normal saline and adjusted to a concentration of 1 in 40 were treated with an equal volume of dilute tannic acid, washed
30
B edi/S ehgal/S engupta/K aur/A ikat
and coated with denatured human y-globulin (63° C for 30 min and diluted to 3%). Hemagglutination tests were put up with doubling dilution of sera and titers noted. (B) Indirect IF technique for detection of nonagglutinating antibodies modified from P eltier et al. [12]. Sheep red cells were treated with an equal volume of 1:200 dilution of amboceptor for 15 min at room temperature. The coated cells were spread on slides and conventional indirect IF technique performed for the detection of nonagglutinating anti-IgG antibodies.
Results The results are depicted in table I. The mean values of serum IgG, IgM, IgA and IgD in 42 patients with psoriasis showed no significant difference from control subjects. With the TRC hemagglutination method, the mean values of the antiglobulin titers in 42 psoriatics and controls were again not significantly different. Indirect IF technique also revealed negative results. The antinuclear factor was weakly positive in sera from only 2 of the 42 patients (4.8%); the positivity rate in healthy population being less than 2% [14].
Comment
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
Our results do not substantiate the previously reported disturbances in serum immunoglobulins in patients with psoriasis (table II). Likewise, we were unable to demonstrate antiglobulin activity in serum reported by G uilhou et al. [4], T apanes et al. [15] also failed to detect such an activity, whereas other workers [2, 4, 5] using immunoadsorption [16] or indirect IF techniques [12] have been successful in demonstrating antiglo bulin factor in sera from patients with psoriasis. Indeed, the results appear to vary depending upon the substrate and the technique used. The negativity of our results raises two possibilities, namely either the serum autoantibodies are not detectable by the technique employed or these are not present in the serum of psoriatic patients. We also employed the same sera in indirect IF tests carried out on normal skin sections and were not able to detect any fluorescence, although our preliminary results from direct IF studies on tissue sections of psoriatic lesions in some of these patients did show focal deposition of immune complexes of IgG, IgM and complement within the layers of the stratum corneum. This prompts us to believe that serum antiglobulin antibodies, perhaps, get drained into the lesion thereby making their detection in the serum
Immunological Studies in Psoriasis
31
Table I. Values of serum immunoglobulins and antiglobulin titers in psoriatic patients and control subjects Serum
IgG, mg% IgM, mg% IgA, mg% IgD, IU Antiglobulin
control psoriasis control psoriasis control psoriasis control psoriasis control psoriasis
Number tested
Mean
SD
p value >
113 42 113 42 20 42 20 42 20 42
1,145.2 1,098.98 144.00 160.69 191.40 169.30 34.60 37.58 14.75
±339.00 ±425.49 ± 60.00 ± 47.73 ± 71.97 ± 56.48 ± 24.57 ± 23.68 ± 10.94 ± 7.31
0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05
11.00
Table II. Serum immunoglobulin data from psoriatic patients compared to previously reported studies in the literature
F raser el al. [3] M arghescu et al. [11] Varelzidis et al. [17] L ai a F at et al. [9] G uilhou et al. [4]
Present study
Number studied
IgG
IgM
IgA
IgD
27 101 105 15 60 42
low high normal low high normal
low normal normal low normal normal
high high high low high normal
not done not done not done not done not done normal
difficult. Further search for autoantibodies or immune complexes both in the serum and at the cellular level using more sensitive techniques may prove helpful in understanding the etiopathogenesis of psoriasis.
References
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
1 B eutner, E.H .; J ablonska, S.; J arsabek-C horzelska, M., et al.: Studies in immunodermatology. VI. Immunofluorescence studies of autoantibodies to the stratum corneum and of in vivo fixed IgG in stratum corneum of psoriatic lesions. Int. Archs Allergy appl. Immun. 48: 301-323 (1975).
32
Bedi/S ehgal/S engupta/K aur/A ikat
2 F lorin-C hristensen, A .; Coceo, M.J.A.; A rana , A.; P orrini, A., et al.: Antigammaglobulin factors in psoriasis. Dermatológica 149: 220-224 (1974). 3 F raser, N.G.; D ick , H.M.; and G richton , W.B.: Immunoglobulins in dermatitis herpetiformis and various other skin diseases. Br. J. Derm. 81: 89-93 (1969). 4 G uilhou , J.; C lot, J.; M eynadier, J., and L apinski, J.: Immunological aspects of psoriasis. I. Immunoglobulins and anti-IgG factors. Br. J. Derm. 94: 501-508 (1976). 5 H owell , F .A .; C hamberlain, M.A.; P erry, R.A., et al.: IgG antiglobulin levels in patients with psoriatic arthropathy, ankylosing spondylitis and gout. Ann. rheum. Dis. 31: 129-131 (1972). 6 J ablonska, S.; C horzelski, T.P.; C horzelska, M.J., and Beutner, E.H.: Studies in immunodermatology. VII. Int. Archs Allergy appl. Immun. 48: 324-340 (1975). 7 K rogh , H.K. and T onder, O.: Immunoglobulins and anti-immunoglobulin factors in psoriatic lesions. Clin. exp. Immunol. 10: 623-634 (1972). g K rogh , H.K. and T onder, O .: Antibodies in psoriatic scales. Scand. J. Immunol. 2: 45-52 (1973). 9 L ai a F at , R .F.M . and Vanfurth , R .: Serum immunoglobulin levels in various skin diseases. Clin. exp. Immunol. 17: 129-137 (1974). 10 M ancini, G .; C arroñara, A.E., and H eremans, J.F .: Immunological quantification of antigens by single radial immunodiffusion. Immunochemistry 2: 235-242 (1965). 11 M arghescu, S. und B raun-F alco , O .: Quantitatives Verhalten von Immunoglobulin und Komplement bei Psoriasis. Arch. klin. exp. Derm. 238:417-420(1970). 12 P eltier, A.; L eroy, C.; A tra , E. et H aim, T .: Detection des facteurs antigammaglobuliniques sériques par immunofluorescence. Path. Biol., Paris 20: 181 (1972). 13 R imbaud, P.; M eynadier, J.; G uilhou , J„ and C lot, J.: Anti IgG activity of peri pheral blood lymphocytes in psoriasis. Archs Derm. 108: 371-373 (1973). ]4 Sehgal, S.; P asricha, N., and A ikat , B.K.: Antinuclear antibodies in health and disease. Ind. J. med. Res. (in press). 15 T apanes, F.J.; R awson, A.J., and H ollander, J.L.: Serum anti-immunoglobulins in psoriatic arthritis as compared with rheumatoid arthritis. Arthritis Rheum. 15:153-156 (1972). 16 T orrigiani, G. and R oitt, I.M .: Antiglobulin factors in sera from patients with rheumatoid arthritis and normal subjects. Ann. rheum. Dis. 26: 334-340 (1967). 17 V erelzidis, A., and T heodoris, A.: Serum immunoglobulins in psoriasis before and after ultraviolet therapy. Br. J. Derm. 85: 14-17 (1971).
T ilak R. B edi, MD, Department of Dermatology, Leprology and STD, Postgraduate Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
Institute of Medical Education and Research, Chandigarh-160011 (India)
Immunological Studies in Psoriasis T.R. Beim, S. Sehgal, S. Sengupta , S. K aur and B.K. A ikat Departments of Dermatology, Immunopathology, and Pathology, Postgraduate Institute of Medical Education and Research, Chandigarh
Key Words. Psoriasis • Immunoglobulins IgG, IgM, IgA, IgD • Antiglobulins Abstract. The mean values of serum immunoglobulins A, G, M and D levels in 42 psoriasis patients showed no significant difference from those of control subjects. Analysis of the same sera for the presence of antiglobulin antibodies also yielded negative results. Antinuclear factor could be demonstrated in only 4.8% of the cases. The possibility that serum antiglobulins get drained into the lesions making their detection in the serum difficult, is supported by our preliminary findings of focal deposition in the stratum corneum of immune complexes of IgG, IgM and complement in tissue sections of psoriatic lesions from these patients.
Received: September 19, 1977; accepted: November 4, 1977.
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
The autoimmune hypothesis regarding etiopathogenesis of psoriasis is largely based on the demonstration of serum antibodies to stratum corneum and complement-mediated antigen-antibody complexes in the superficial layers of the psoriatic epidermis [1, 6-8], The immune complexes, perhaps, initiate a chain of events eliciting chemotactic cellular reaction resulting in the histogenesis of psoriatic lesion. Antiglobulin factors have been shown in the psoriatic lesion [2, 5] and studies on serum immunoglobulins have yielded rather contradictory results. Variously, the serum IgG has been reported to be normal [17], low [3, 9] or high [4, 11]. Serum IgM is reported to be either normal [4, 11, 17] or low [3, 9], Apart from a solitary report of low serum IgA [9], it has been shown by many workers to be raised [3, 4, 11, 17], Results of serum IgD estimations in psoriasis are not available.
Immunological Studies in Psoriasis
29
Anti-IgG activity has been shown to be present on blood lymphocytes from patients with psoriasis [13]. The success to demonstrate antiglobulin factors in the serum, however, appears to be related to the specificity of anti globulin and the technique employed for its detection [2], We have attempted to evaluate serum immunoglobulins G, M, A and D by conventional methods and antiglobulin factors in the serum employing techniques, both for agglutinating and nonagglutinating antibodies. The results are presented herein.
Materials and Methods The investigation was carried out on 42 patients (12 female and 30 male) with psoriasis; the age ranging from 15 to 55 years and the duration of the disease varying from 2 months to 30 years. The body surface area affected varied from 20 to 100%; arthralgia was present in 6 patients including 1 having deforming arthropathy of the hands and feet. 37 patients had received topical tar applications; 6 of them also received topical corticosteroid treatment at some time during their illness and the other 5 had no treatment prior to the study. None had received internal corticosteroids or cytotoxic drugs. The control group comprised 20 age-matched normal healthy individuals. Serum Immunoglobulin Estimation The quantitative determination of serum IgG, IgM, IgA and IgD were carried out by the radial immunodiffusion technique [10] using monospecific antisera obtained from Hyland Laboratory, USA. Reference standard serum for immunoglobulins was obtained from WHO (courtesy D.S. R owe). Antinuclear Antibodies The conventional indirect IF technique was employed. Briefly 4 ¡xm thick air-dried cryostat sections of rat liver were layered with 1:10 dilution patients’ serum for 30 min at 37 °C, washed in several changes of phosphate-buffered saline (pH 7.4) and treated with fluorescein-labelled polyvalent antihuman immunoglobulin antiserum (Burroughs-Wellcome) at a dilution of 1 in 10 for another 30 min at 37°C. The sections were washed in phosphate-buffered saline and mounted in buffered glycerin and examined under fluores cent light.
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
Antiglobulin Factor Antiglobulin factors were searched for in the patients sera employing 2 different tech niques: (A) Tanned red cell (TRC) hemagglutination method employing denatured human y-globulin (Curewell India Ltd.). Sheep red cells collected in Alsevers solution, washed in normal saline and adjusted to a concentration of 1 in 40 were treated with an equal volume of dilute tannic acid, washed
30
B edi/S ehgal/S engupta/K aur/A ikat
and coated with denatured human y-globulin (63° C for 30 min and diluted to 3%). Hemagglutination tests were put up with doubling dilution of sera and titers noted. (B) Indirect IF technique for detection of nonagglutinating antibodies modified from P eltier et al. [12]. Sheep red cells were treated with an equal volume of 1:200 dilution of amboceptor for 15 min at room temperature. The coated cells were spread on slides and conventional indirect IF technique performed for the detection of nonagglutinating anti-IgG antibodies.
Results The results are depicted in table I. The mean values of serum IgG, IgM, IgA and IgD in 42 patients with psoriasis showed no significant difference from control subjects. With the TRC hemagglutination method, the mean values of the antiglobulin titers in 42 psoriatics and controls were again not significantly different. Indirect IF technique also revealed negative results. The antinuclear factor was weakly positive in sera from only 2 of the 42 patients (4.8%); the positivity rate in healthy population being less than 2% [14].
Comment
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
Our results do not substantiate the previously reported disturbances in serum immunoglobulins in patients with psoriasis (table II). Likewise, we were unable to demonstrate antiglobulin activity in serum reported by G uilhou et al. [4], T apanes et al. [15] also failed to detect such an activity, whereas other workers [2, 4, 5] using immunoadsorption [16] or indirect IF techniques [12] have been successful in demonstrating antiglo bulin factor in sera from patients with psoriasis. Indeed, the results appear to vary depending upon the substrate and the technique used. The negativity of our results raises two possibilities, namely either the serum autoantibodies are not detectable by the technique employed or these are not present in the serum of psoriatic patients. We also employed the same sera in indirect IF tests carried out on normal skin sections and were not able to detect any fluorescence, although our preliminary results from direct IF studies on tissue sections of psoriatic lesions in some of these patients did show focal deposition of immune complexes of IgG, IgM and complement within the layers of the stratum corneum. This prompts us to believe that serum antiglobulin antibodies, perhaps, get drained into the lesion thereby making their detection in the serum
Immunological Studies in Psoriasis
31
Table I. Values of serum immunoglobulins and antiglobulin titers in psoriatic patients and control subjects Serum
IgG, mg% IgM, mg% IgA, mg% IgD, IU Antiglobulin
control psoriasis control psoriasis control psoriasis control psoriasis control psoriasis
Number tested
Mean
SD
p value >
113 42 113 42 20 42 20 42 20 42
1,145.2 1,098.98 144.00 160.69 191.40 169.30 34.60 37.58 14.75
±339.00 ±425.49 ± 60.00 ± 47.73 ± 71.97 ± 56.48 ± 24.57 ± 23.68 ± 10.94 ± 7.31
0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05 0.05
11.00
Table II. Serum immunoglobulin data from psoriatic patients compared to previously reported studies in the literature
F raser el al. [3] M arghescu et al. [11] Varelzidis et al. [17] L ai a F at et al. [9] G uilhou et al. [4]
Present study
Number studied
IgG
IgM
IgA
IgD
27 101 105 15 60 42
low high normal low high normal
low normal normal low normal normal
high high high low high normal
not done not done not done not done not done normal
difficult. Further search for autoantibodies or immune complexes both in the serum and at the cellular level using more sensitive techniques may prove helpful in understanding the etiopathogenesis of psoriasis.
References
Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
1 B eutner, E.H .; J ablonska, S.; J arsabek-C horzelska, M., et al.: Studies in immunodermatology. VI. Immunofluorescence studies of autoantibodies to the stratum corneum and of in vivo fixed IgG in stratum corneum of psoriatic lesions. Int. Archs Allergy appl. Immun. 48: 301-323 (1975).
32
Bedi/S ehgal/S engupta/K aur/A ikat
2 F lorin-C hristensen, A .; Coceo, M.J.A.; A rana , A.; P orrini, A., et al.: Antigammaglobulin factors in psoriasis. Dermatológica 149: 220-224 (1974). 3 F raser, N.G.; D ick , H.M.; and G richton , W.B.: Immunoglobulins in dermatitis herpetiformis and various other skin diseases. Br. J. Derm. 81: 89-93 (1969). 4 G uilhou , J.; C lot, J.; M eynadier, J., and L apinski, J.: Immunological aspects of psoriasis. I. Immunoglobulins and anti-IgG factors. Br. J. Derm. 94: 501-508 (1976). 5 H owell , F .A .; C hamberlain, M.A.; P erry, R.A., et al.: IgG antiglobulin levels in patients with psoriatic arthropathy, ankylosing spondylitis and gout. Ann. rheum. Dis. 31: 129-131 (1972). 6 J ablonska, S.; C horzelski, T.P.; C horzelska, M.J., and Beutner, E.H.: Studies in immunodermatology. VII. Int. Archs Allergy appl. Immun. 48: 324-340 (1975). 7 K rogh , H.K. and T onder, O.: Immunoglobulins and anti-immunoglobulin factors in psoriatic lesions. Clin. exp. Immunol. 10: 623-634 (1972). g K rogh , H.K. and T onder, O .: Antibodies in psoriatic scales. Scand. J. Immunol. 2: 45-52 (1973). 9 L ai a F at , R .F.M . and Vanfurth , R .: Serum immunoglobulin levels in various skin diseases. Clin. exp. Immunol. 17: 129-137 (1974). 10 M ancini, G .; C arroñara, A.E., and H eremans, J.F .: Immunological quantification of antigens by single radial immunodiffusion. Immunochemistry 2: 235-242 (1965). 11 M arghescu, S. und B raun-F alco , O .: Quantitatives Verhalten von Immunoglobulin und Komplement bei Psoriasis. Arch. klin. exp. Derm. 238:417-420(1970). 12 P eltier, A.; L eroy, C.; A tra , E. et H aim, T .: Detection des facteurs antigammaglobuliniques sériques par immunofluorescence. Path. Biol., Paris 20: 181 (1972). 13 R imbaud, P.; M eynadier, J.; G uilhou , J„ and C lot, J.: Anti IgG activity of peri pheral blood lymphocytes in psoriasis. Archs Derm. 108: 371-373 (1973). ]4 Sehgal, S.; P asricha, N., and A ikat , B.K.: Antinuclear antibodies in health and disease. Ind. J. med. Res. (in press). 15 T apanes, F.J.; R awson, A.J., and H ollander, J.L.: Serum anti-immunoglobulins in psoriatic arthritis as compared with rheumatoid arthritis. Arthritis Rheum. 15:153-156 (1972). 16 T orrigiani, G. and R oitt, I.M .: Antiglobulin factors in sera from patients with rheumatoid arthritis and normal subjects. Ann. rheum. Dis. 26: 334-340 (1967). 17 V erelzidis, A., and T heodoris, A.: Serum immunoglobulins in psoriasis before and after ultraviolet therapy. Br. J. Derm. 85: 14-17 (1971).
T ilak R. B edi, MD, Department of Dermatology, Leprology and STD, Postgraduate Downloaded by: King's College London 137.73.144.138 - 3/7/2018 7:26:45 AM
Institute of Medical Education and Research, Chandigarh-160011 (India)
