Clin. exp. Immunol. (1977) 29, 23-29.
Circulating immune complexes in retinal vasculitis B. S. ANDREWS, JILL MCINTOSH, VALERIE PETTS & R. PENNY Department of Immunology, St Vincent's Hospital, and the Department of Medicine, University of New South Wales, Sydney, Australia
(Received 5 January 1977)
SUMMARY
Seventeen patients with retinal vasculitis, eleven with the peripheral type (Eales' disease) and six with the central type, were investigated to detect the presence of circulating immune complexes (IC) which might then be related to the pathogenesis of their disease. A systemic disease process was identified in six. IC in serum were inferred by the presence of complement (C) activation, rheumatoid factor, Clq or monoclonal rheumatoid factor precipitins, anticomplementary activity, elevated cryoglobulins, inhibition of erythrocyte-antibody (IgG-EA) rosette formation, increased numbers of peripheral blood lymphocytes bearing surface Ig, and spontaneous neutrophil chemotactic activity in plasma. Two or more parameters were positive in thirteen of seventeen patients, with chemotactic activity (69%) and inhibition of EA-rosette formation (590%) being the most frequently positive tests. No immunological differences were detected between the peripheral and central retinalvasculitic groups. Several IC systems may operate in a given patient.
INTRODUCTION Retinal vasculitis (RV) is an inflammatory process involving predominantly the retinal venous system, although it may occasionally extend to involve the arterial system. The cause is unknown for the peripheral type of RV (PRV), described originally by Henry Eales (Eales & Birmingham, 1880), and found frequently in young adults in association with intraocular haemorrhage and variable loss of vision. Central retinal vasculitis (CRV), an entity first described in 1961 by Lyle & Wybar is regarded as a benign, unilateral disease with a male predominance which, although protracted, only occasionally results in impairment of visual acuity. Hart et al. (1971) believe the entity of CRV to be the result of a central vein occlusion in an otherwise normal retinal vascular tree, although the occurrence of CRV in one eye and PRV in the other (Lyle & Wybar, 1961), would suggest that CRV may have an inflammatory origin. However, experimental evidence showed that an immunological reaction may be involved, mediated by local deposition of IC. De Muro & Focosi (1951), and, more recently, Levine & Ward (1970) injected antigen (Ag) into the vitreous humour of sensitized animals which produced a local Arthus-type reaction. Ag and antibody (Ab) were demonstrated in the retinal vessel walls and persisted for up to 8 days. In addition, the occasional association of RV with systemic inflammatory disease suggested that RV may be due to deposition of circulating Ag-Ab complexes. With this hypothesis, the present study was undertaken to determine if immune complexes (IC) were present in serum and if so, what role they may play in the basic immunopathology of the disease.
MATERIALS AND METHODS Patient selection. Patients with a diagnosis of peripheral or central retinal vasculitis were referred to the Immunology Unit, St Vincent's Hospital, Sydney, by ophthalmologists at the Eye Department, St Vincent's Hospital, the Prince of Wales
Correspondence: Professor R. Penny, Department of Immunology, St Vincent's Hospital, Darlinghurst 2010 N.S.W., Australia.
23
24
B. S. Andrews et al.
Hospital and the Sydney Eye Hospital. The diagnosis of CRV was only made in the absence of hypertension, degenerativ e retinal arterial disease and raised intracranial pressure. Patients were routinely seen in consultation with a neurologist. Fluorescein angiography was consistent with the diagnosis in all patients. Patients with systemic lupus erythematosus were not included in the study. Immunological investigations. Serum levels of IgG, IgA and IgM and plasma C3 and C4 levels were quantitated by commercial radial immunodiffusion plates (Hyland Laboratories, Costa Mesa, California). Rheumatoid factor (RF) in serum was detected by IgG-coated latex particles (Hyland Laboratories, Costa Mesa, California) and titred using the sensitized sheep red cell technique (Wellcome Diagnostics, Sydney, Australia). Conversion of C3 to C3c was determined by Ag-Ab twodimensional crossed electrophoresis. Membrane-bound immunoglobulin (SmIg) on peripheral blood lymphocytes (PBL) was performed as described by Cooper et al. (1975), increased levels implying adsorbed IC onto Fc receptors of B lymphocytes. The anticomplementary assay (AC) was carried out as described by Johnson, Mowbray & Porter (1975) and the results expressed as the per cent inhibition of haemolysis of sensitized sheep red cells. Cryoglobulins in serum were detected using slight modifications to the described methods (Cream, 1971). Serum, collected in a pre-warmed 'Vacutainer' syringe, clotted and separated at 37 C, was allowed to stand for 5 days at 4VC. After centrifugation at 4VC the precipitate was washed four times in phosphate-buffered saline (PBS), pH 7 2, at 4VC, resuspended in 2 ml PBS and warmed at 37 C for 4 hr. After centrifugation at 37 C to remove insoluble material, the supernatant was separated, its protein content estimated by the Lowry technique and corrected for the original serum volume. Semiquantitative assessment of the IgG, IgA, IgM, Clq, C3 and C4 was made by two-dimensional double immunodiffusion. Clq was isolated according to Agnello, Winchester & Kunkel (1970) and used at a concentration of 0-2 mg/ml in gel diffusion. Highly purified monoclonal K type IgM with RF actixity (1:4096) was isolated (Andrews et al., 1976; Winchester, Kunkel & Agnello, 1971) from a mixed cryoglobulin and was employed at a concentration of 3 4 mg/ml in gel diffusion as described (Winchester et al., 1971). EA-rosette inhibition. This method has been described in detail elsewhere (Andrews et al., 1977), but, in essence, PBL were isolated on Ficoll-Hypaque density gradient, washed three times in PBS and reconstituted to 0 5 x 106 cells in 100,ul Hanks' BSS containing 10% heat-inactivated and red cell-absorbed foetal calf serum (Hanks'-FCS). Duplicate 100 Pl PBL suspensions were incubated with 100 P1 of decomplemented test serum for 30 min at 37°C. Cells were centrifuged, washed three times in PBS at 4°C, then 200,ul optimally sensitized IgG-EA sheep red cells (EA-SRBC) (0-500 v/v) were added to the pellet. After resuspending, the mixture was incubated at 37°C for 10 min, the cells pelleted at 50 g for 5 min and further incubated at 37°C for 30 min. After brisk xortexing, the number of PBLs rosetting four or more EA-SRBC was determined. The per cent inhibition of EA rosettes formed with test serum, compared with the control of cells incubated in Hanks'-FCS, was calculated. Using trypan blue the viability of lymphocytes was assessed to be in excess of 950%,. Neutrophil chemotactic activity in plasma. This was measured using a modification of the Boyden Chamber method, previously described in detail (Andrews et al., 1977). Chemotaxis was determined by counting the number of neutrophils migrating to the lower surface of a 3,um 'Sartorius' membrane after 2 5 hr incubation at 37°C, 2x 106 neutrophils from group 0 donors were placed in the upper chamber and 500 heparinized test plasma in Hanks' BSS, in the lower chamber. The chemotactic index (CI) was calculated by counting the number of neutrophils on six-high power fields of the attractant sides of each of three membranes, taking a mean and subtracting from this mean that produced using plasma from normal AB group donors. Controls using casein, aggregated IgG and 75 IgG as described previously (Andrews et al., 1977) were used. Statistical analyses were performed on groups by the Wilcoxon-Rank sum test, Mann-Whitney U test and the squared analysis using 2 x 2 contingency tables. Significance was assessed below the 50 0 lex el.
RESULTS Clinical data Table 1 shows the sex of the patients, the age at onset, and duration of disease: this extended from 6-48 months in PRV and 6-24 months in CVR. Bilateral inflammatory disease was present in 82% of patients uxith PRV and in none with CRV. Associated mild anterior uveitis was present in four PRV and one CRY, with posterior uveitis present in one patient in each group. Visual acuity (VA), related to the inflammatory process and retinal and vitreous haemorrhage, was reduced to 6/18 or less, in 7300 of patients with PRV: in 830/ of patients with CRV, VA xas invariably associated with marked macular oedema. Systemic corticosteroids were used in all patients except case No. 16, with improvement in VA noted in four patients with PRV, and in one with CRV. Obvious cessation of the total inflammatory disease process was seen in two patients (cases 7 and 8) with Behcet's syndrome. Glomerulonephritis in two patients was confirmed by renal biospy, there was one diffuse proliferative type (case 5) and one focal proliferative type (case 16); Behcet's syndrome was found in cases 7 and 8; brain stem vasculitis in case 3; and Cushing's disease in case 13 associated with rheumatoid arthritis (RA) and IgA deficiency. Case 11
Immune complexes in retinal vasculitis
25
TABLE 1. Clinical details of seventeen patients with retinal vasculitis*
Case No.
Sex
onset
Duration (months)
1 2 3 4 5 6 7 8 9 10 11 12t 13 14t 15 16 17
F F M F M F F F F F M F M M M M F
31 26 25 52 48 43 33 27 50 30 42 49 41 44 50 42 61
21 30 48 27 8 24 24 21 6 36 9 24 18 15 15 12 6
Age
*
Bilateral
Associated uveitis
+ + + + + + + + +
+ (Ant) + (Ant) + (Ant & Post) -
-
-
+ (Ant)
Associated conditions
Brain stem vasculitis
Glomcrulonephritis, diabetes Behcet's syndrome Behcet's syndrome
-
-
+ (Ant & Post) -
Nasopharyngeal cancer/DXRT
Cushing's disease, RA, IgA deficiency Glomerulonephritis
1-11 Peripheral type, 12-17 central type.
t Radiological evidence (asymptomatic) of apical dental-filling defect.
developed unilateral PRV 18 months after surgical removal of a nasopharyngeal carcinoma with followup radiotherapy. Plasma proteins, serology Serum Ig levels were normal in eleven patients. Case 13 had IgA deficiency while five patients had elevated IgA and/or IgM levels. No significant abnormalities were present on serum protein electrophoresis. Antinuclear antibodies above a titre of 1:10 were not detected. No laboratory evidence of streptococcal infection, toxoplasmosis, histoplasmosis, syphilis or recent ocular trauma was found. Elevated plasma cryofibrinogen was present in six patients (5, 8-11, 16), three of whom were receiving oestrogens.
Assays for circulating immune complexes As shown in Table 2, four patients had serum RF detected in titres ranging from 1:8-1:32 by the sheep red cell test (the normal for our laboratory was < 1: 8). Complement activation was indicated in four patients: there was a reduction in C3 level (normal 80-120 mg0%O) in case 5; a reduction in C3, C4 (normal 20-40 mg%O) and C3 conversion to C3c in case 4; and isolated partial conversion to C3c as the only abnormality was found in cases 3 and 12. No precipitins were detected in any case using purified Clq or monoclonal RF. SmIg-positive PBL were slightly increased in cases 6 and 11. Normal laboratory ranges from SmIgG (0-295 cells/ul), SmIgA (0-100 cells/ul) and SmIgM (0-207 cells/gl) have been described elsewhere (Cooper et al., 1975). Case 6 had 390 SmIgG-positive cells/pl and 234 SmIgM-positive cells/,ul, while case 11 had 235 SmIgM-positive cells/ul. Thirty-five normal control sera showed no AC activity (Fig. 1) and zero inhibition of haemolysis. Four of sixteen patients (250%) had AC activity detected in serum (Table 2) with a mean inhibition of 10.4% with 2 s.d. of 42-8%. A significant difference between the groups was detected using x2 analysis (P,2 24
24
25
25
69
25
59
positive
77% 'X No precipitins were detected sN ith Clq or mRF. 1OO
90_ n 16 =
S
300H
70-
n- 16 0
,>
0 ID
60[ .
0 -C
50 _
200k-
c
E
40_
a) 0
30_
S
n=33
100
20k_
0%
1o0-
0@ 0O
0
tS
n=35
0
I'VLWP
z
Controls
Retinal
C on trolIs
Retinal
vascu litis
vasculitis
FIG. 2
FIG.
FiG. 1. Anti-complementary activity in
serum
of sixteen
patients
with retinal vasculitis. Horizontal bars
represent mean for each group. FiG. 2. Mixed cryoglobulins in serum of sixteen patients with retinal vasculitis. Lower and upper horizontal bars in control group represent mean and mean -t2 s.d. respectively. Horizontal bar in retinal vasculitis group represents mean.
Immune complexes in retinal vasculitis
70r
27
200k n=17
60k-
0
50k-
0
40k
0 B 0
n=40 -c
:L
30k
0
150
0 -
n' 16
0@0 0
~~0-~
0
0
0
c
0
0
E
look
0 a) _C
0
0@0
0
0@0
0
20k
00 00 0
0
0% 10
0
0@ n =34
0
0.
0
50k
0 0
0
00000000
Controls
Retinal vasculitis FIG. 3
OL
0
l& Con trols
Retinal vasculitis FIG. 4
FIG. 3. Inhibition of EA-rosette formation by serum of seventeen patients with retinal vasculitis. For explanation of horizontal bars see Fig. 2. FIG. 4. Neutrophil chemotactic activity of plasma from 16 patients with retinal vasculitis. For explanation of horizontal bars see Fig. 2.
neutrophil CI, a significant direct correlation with the cryoglobulin level was found (r = 0-828, P
Circulating immune complexes in retinal vasculitis B. S. ANDREWS, JILL MCINTOSH, VALERIE PETTS & R. PENNY Department of Immunology, St Vincent's Hospital, and the Department of Medicine, University of New South Wales, Sydney, Australia
(Received 5 January 1977)
SUMMARY
Seventeen patients with retinal vasculitis, eleven with the peripheral type (Eales' disease) and six with the central type, were investigated to detect the presence of circulating immune complexes (IC) which might then be related to the pathogenesis of their disease. A systemic disease process was identified in six. IC in serum were inferred by the presence of complement (C) activation, rheumatoid factor, Clq or monoclonal rheumatoid factor precipitins, anticomplementary activity, elevated cryoglobulins, inhibition of erythrocyte-antibody (IgG-EA) rosette formation, increased numbers of peripheral blood lymphocytes bearing surface Ig, and spontaneous neutrophil chemotactic activity in plasma. Two or more parameters were positive in thirteen of seventeen patients, with chemotactic activity (69%) and inhibition of EA-rosette formation (590%) being the most frequently positive tests. No immunological differences were detected between the peripheral and central retinalvasculitic groups. Several IC systems may operate in a given patient.
INTRODUCTION Retinal vasculitis (RV) is an inflammatory process involving predominantly the retinal venous system, although it may occasionally extend to involve the arterial system. The cause is unknown for the peripheral type of RV (PRV), described originally by Henry Eales (Eales & Birmingham, 1880), and found frequently in young adults in association with intraocular haemorrhage and variable loss of vision. Central retinal vasculitis (CRV), an entity first described in 1961 by Lyle & Wybar is regarded as a benign, unilateral disease with a male predominance which, although protracted, only occasionally results in impairment of visual acuity. Hart et al. (1971) believe the entity of CRV to be the result of a central vein occlusion in an otherwise normal retinal vascular tree, although the occurrence of CRV in one eye and PRV in the other (Lyle & Wybar, 1961), would suggest that CRV may have an inflammatory origin. However, experimental evidence showed that an immunological reaction may be involved, mediated by local deposition of IC. De Muro & Focosi (1951), and, more recently, Levine & Ward (1970) injected antigen (Ag) into the vitreous humour of sensitized animals which produced a local Arthus-type reaction. Ag and antibody (Ab) were demonstrated in the retinal vessel walls and persisted for up to 8 days. In addition, the occasional association of RV with systemic inflammatory disease suggested that RV may be due to deposition of circulating Ag-Ab complexes. With this hypothesis, the present study was undertaken to determine if immune complexes (IC) were present in serum and if so, what role they may play in the basic immunopathology of the disease.
MATERIALS AND METHODS Patient selection. Patients with a diagnosis of peripheral or central retinal vasculitis were referred to the Immunology Unit, St Vincent's Hospital, Sydney, by ophthalmologists at the Eye Department, St Vincent's Hospital, the Prince of Wales
Correspondence: Professor R. Penny, Department of Immunology, St Vincent's Hospital, Darlinghurst 2010 N.S.W., Australia.
23
24
B. S. Andrews et al.
Hospital and the Sydney Eye Hospital. The diagnosis of CRV was only made in the absence of hypertension, degenerativ e retinal arterial disease and raised intracranial pressure. Patients were routinely seen in consultation with a neurologist. Fluorescein angiography was consistent with the diagnosis in all patients. Patients with systemic lupus erythematosus were not included in the study. Immunological investigations. Serum levels of IgG, IgA and IgM and plasma C3 and C4 levels were quantitated by commercial radial immunodiffusion plates (Hyland Laboratories, Costa Mesa, California). Rheumatoid factor (RF) in serum was detected by IgG-coated latex particles (Hyland Laboratories, Costa Mesa, California) and titred using the sensitized sheep red cell technique (Wellcome Diagnostics, Sydney, Australia). Conversion of C3 to C3c was determined by Ag-Ab twodimensional crossed electrophoresis. Membrane-bound immunoglobulin (SmIg) on peripheral blood lymphocytes (PBL) was performed as described by Cooper et al. (1975), increased levels implying adsorbed IC onto Fc receptors of B lymphocytes. The anticomplementary assay (AC) was carried out as described by Johnson, Mowbray & Porter (1975) and the results expressed as the per cent inhibition of haemolysis of sensitized sheep red cells. Cryoglobulins in serum were detected using slight modifications to the described methods (Cream, 1971). Serum, collected in a pre-warmed 'Vacutainer' syringe, clotted and separated at 37 C, was allowed to stand for 5 days at 4VC. After centrifugation at 4VC the precipitate was washed four times in phosphate-buffered saline (PBS), pH 7 2, at 4VC, resuspended in 2 ml PBS and warmed at 37 C for 4 hr. After centrifugation at 37 C to remove insoluble material, the supernatant was separated, its protein content estimated by the Lowry technique and corrected for the original serum volume. Semiquantitative assessment of the IgG, IgA, IgM, Clq, C3 and C4 was made by two-dimensional double immunodiffusion. Clq was isolated according to Agnello, Winchester & Kunkel (1970) and used at a concentration of 0-2 mg/ml in gel diffusion. Highly purified monoclonal K type IgM with RF actixity (1:4096) was isolated (Andrews et al., 1976; Winchester, Kunkel & Agnello, 1971) from a mixed cryoglobulin and was employed at a concentration of 3 4 mg/ml in gel diffusion as described (Winchester et al., 1971). EA-rosette inhibition. This method has been described in detail elsewhere (Andrews et al., 1977), but, in essence, PBL were isolated on Ficoll-Hypaque density gradient, washed three times in PBS and reconstituted to 0 5 x 106 cells in 100,ul Hanks' BSS containing 10% heat-inactivated and red cell-absorbed foetal calf serum (Hanks'-FCS). Duplicate 100 Pl PBL suspensions were incubated with 100 P1 of decomplemented test serum for 30 min at 37°C. Cells were centrifuged, washed three times in PBS at 4°C, then 200,ul optimally sensitized IgG-EA sheep red cells (EA-SRBC) (0-500 v/v) were added to the pellet. After resuspending, the mixture was incubated at 37°C for 10 min, the cells pelleted at 50 g for 5 min and further incubated at 37°C for 30 min. After brisk xortexing, the number of PBLs rosetting four or more EA-SRBC was determined. The per cent inhibition of EA rosettes formed with test serum, compared with the control of cells incubated in Hanks'-FCS, was calculated. Using trypan blue the viability of lymphocytes was assessed to be in excess of 950%,. Neutrophil chemotactic activity in plasma. This was measured using a modification of the Boyden Chamber method, previously described in detail (Andrews et al., 1977). Chemotaxis was determined by counting the number of neutrophils migrating to the lower surface of a 3,um 'Sartorius' membrane after 2 5 hr incubation at 37°C, 2x 106 neutrophils from group 0 donors were placed in the upper chamber and 500 heparinized test plasma in Hanks' BSS, in the lower chamber. The chemotactic index (CI) was calculated by counting the number of neutrophils on six-high power fields of the attractant sides of each of three membranes, taking a mean and subtracting from this mean that produced using plasma from normal AB group donors. Controls using casein, aggregated IgG and 75 IgG as described previously (Andrews et al., 1977) were used. Statistical analyses were performed on groups by the Wilcoxon-Rank sum test, Mann-Whitney U test and the squared analysis using 2 x 2 contingency tables. Significance was assessed below the 50 0 lex el.
RESULTS Clinical data Table 1 shows the sex of the patients, the age at onset, and duration of disease: this extended from 6-48 months in PRV and 6-24 months in CVR. Bilateral inflammatory disease was present in 82% of patients uxith PRV and in none with CRV. Associated mild anterior uveitis was present in four PRV and one CRY, with posterior uveitis present in one patient in each group. Visual acuity (VA), related to the inflammatory process and retinal and vitreous haemorrhage, was reduced to 6/18 or less, in 7300 of patients with PRV: in 830/ of patients with CRV, VA xas invariably associated with marked macular oedema. Systemic corticosteroids were used in all patients except case No. 16, with improvement in VA noted in four patients with PRV, and in one with CRV. Obvious cessation of the total inflammatory disease process was seen in two patients (cases 7 and 8) with Behcet's syndrome. Glomerulonephritis in two patients was confirmed by renal biospy, there was one diffuse proliferative type (case 5) and one focal proliferative type (case 16); Behcet's syndrome was found in cases 7 and 8; brain stem vasculitis in case 3; and Cushing's disease in case 13 associated with rheumatoid arthritis (RA) and IgA deficiency. Case 11
Immune complexes in retinal vasculitis
25
TABLE 1. Clinical details of seventeen patients with retinal vasculitis*
Case No.
Sex
onset
Duration (months)
1 2 3 4 5 6 7 8 9 10 11 12t 13 14t 15 16 17
F F M F M F F F F F M F M M M M F
31 26 25 52 48 43 33 27 50 30 42 49 41 44 50 42 61
21 30 48 27 8 24 24 21 6 36 9 24 18 15 15 12 6
Age
*
Bilateral
Associated uveitis
+ + + + + + + + +
+ (Ant) + (Ant) + (Ant & Post) -
-
-
+ (Ant)
Associated conditions
Brain stem vasculitis
Glomcrulonephritis, diabetes Behcet's syndrome Behcet's syndrome
-
-
+ (Ant & Post) -
Nasopharyngeal cancer/DXRT
Cushing's disease, RA, IgA deficiency Glomerulonephritis
1-11 Peripheral type, 12-17 central type.
t Radiological evidence (asymptomatic) of apical dental-filling defect.
developed unilateral PRV 18 months after surgical removal of a nasopharyngeal carcinoma with followup radiotherapy. Plasma proteins, serology Serum Ig levels were normal in eleven patients. Case 13 had IgA deficiency while five patients had elevated IgA and/or IgM levels. No significant abnormalities were present on serum protein electrophoresis. Antinuclear antibodies above a titre of 1:10 were not detected. No laboratory evidence of streptococcal infection, toxoplasmosis, histoplasmosis, syphilis or recent ocular trauma was found. Elevated plasma cryofibrinogen was present in six patients (5, 8-11, 16), three of whom were receiving oestrogens.
Assays for circulating immune complexes As shown in Table 2, four patients had serum RF detected in titres ranging from 1:8-1:32 by the sheep red cell test (the normal for our laboratory was < 1: 8). Complement activation was indicated in four patients: there was a reduction in C3 level (normal 80-120 mg0%O) in case 5; a reduction in C3, C4 (normal 20-40 mg%O) and C3 conversion to C3c in case 4; and isolated partial conversion to C3c as the only abnormality was found in cases 3 and 12. No precipitins were detected in any case using purified Clq or monoclonal RF. SmIg-positive PBL were slightly increased in cases 6 and 11. Normal laboratory ranges from SmIgG (0-295 cells/ul), SmIgA (0-100 cells/ul) and SmIgM (0-207 cells/gl) have been described elsewhere (Cooper et al., 1975). Case 6 had 390 SmIgG-positive cells/pl and 234 SmIgM-positive cells/,ul, while case 11 had 235 SmIgM-positive cells/ul. Thirty-five normal control sera showed no AC activity (Fig. 1) and zero inhibition of haemolysis. Four of sixteen patients (250%) had AC activity detected in serum (Table 2) with a mean inhibition of 10.4% with 2 s.d. of 42-8%. A significant difference between the groups was detected using x2 analysis (P,2 24
24
25
25
69
25
59
positive
77% 'X No precipitins were detected sN ith Clq or mRF. 1OO
90_ n 16 =
S
300H
70-
n- 16 0
,>
0 ID
60[ .
0 -C
50 _
200k-
c
E
40_
a) 0
30_
S
n=33
100
20k_
0%
1o0-
0@ 0O
0
tS
n=35
0
I'VLWP
z
Controls
Retinal
C on trolIs
Retinal
vascu litis
vasculitis
FIG. 2
FIG.
FiG. 1. Anti-complementary activity in
serum
of sixteen
patients
with retinal vasculitis. Horizontal bars
represent mean for each group. FiG. 2. Mixed cryoglobulins in serum of sixteen patients with retinal vasculitis. Lower and upper horizontal bars in control group represent mean and mean -t2 s.d. respectively. Horizontal bar in retinal vasculitis group represents mean.
Immune complexes in retinal vasculitis
70r
27
200k n=17
60k-
0
50k-
0
40k
0 B 0
n=40 -c
:L
30k
0
150
0 -
n' 16
0@0 0
~~0-~
0
0
0
c
0
0
E
look
0 a) _C
0
0@0
0
0@0
0
20k
00 00 0
0
0% 10
0
0@ n =34
0
0.
0
50k
0 0
0
00000000
Controls
Retinal vasculitis FIG. 3
OL
0
l& Con trols
Retinal vasculitis FIG. 4
FIG. 3. Inhibition of EA-rosette formation by serum of seventeen patients with retinal vasculitis. For explanation of horizontal bars see Fig. 2. FIG. 4. Neutrophil chemotactic activity of plasma from 16 patients with retinal vasculitis. For explanation of horizontal bars see Fig. 2.
neutrophil CI, a significant direct correlation with the cryoglobulin level was found (r = 0-828, P
