Scand J Rheumatology 4: 165-168, 1975
MYCOPLASMA ANTIBODIES IN RHEUMATOID ARTHRITIS Elli Jansson, Paavo Makisara and Sirkka Tuuri
Scand J Rheumatol Downloaded from informahealthcare.com by Cornell University on 12/28/14 For personal use only.
From the University of Tampere, School of Medicine, 33520 Tampere, and Rheumatism Foundation Hospital, 18120 Heinola, Finland
ABSTRACT. Sera of 100 patients under examination at the Outpatient Department of the Rheumatism Foundation Hospital were studied by the indirect hemagglutination technique, using both mycoplasma reference strains, and isolates from RA and SLE as antigens. The series consisted of five groups: I, definite RA (49 patients); 11, probable RA (11);111, possible RA or nonspecific inflammatory arthritis (34); IV, osteoarthrosis (2); V, Reiter's disease (4). Mycoplasma antibodies in titres of 16 or higher were encountered in groups I-IV in 26,8,19, and one case respectively. Twentyone out of 106 blood donors had antibodies against an isolate from RA and/or M. urthn'tidis strain PG 6. The titres found were 16 or 32, except in two cases, 128. In the definite RA group, 21 out of 26 patients possessing mycoplasma antibodies, showed titres of 16 or higher against isolates from RA and/or SLE, 12 against M. urthritidis strain PG 6 and/or Campo, 8 against M. fermentuns, and 6 against a T-strain from NGU. Antibodies against M. arthn'tidis strain Campo were found more often than against strain PG 6. The longer the duration of the arthritic symptoms was, the more frequent seemed also to be the occurrence of mycoplasma antibodies.
Antibodies against Mycoplasma arthritidis (MA) strain Campo, M. fermentans and a f e w isolates from human rheumatoid arthritis (RA) were investigated earlier in sera from patients with this disease. Complement-fixing or metabolism-inhibiting antibodies were not usually detected (2). However, low antibody titres against a human isolate (strain 176-M) were observed by the indirect hemagglutination (IHA) technique. Out of 421 sera with a Waaler-Rose titre of 2128, 30% possessed antibodies in a titre of 2 8 , a n d 19% in a titre of 216
M A T E R I A L AND METHODS Sera These were collected from patients under examination at the Outpatient Department of the Rheumatism Foundation Hospital, Heinola. The series is not entirely unselected as the proportion of relatively new and obscure cases is accented. Patients fulfilling at least five ARA criteria were classified as definite RA, those with a minimum of three as probable RA. The RA group was also subdivided by ARA criteria concerning anatomical joint changes (7). The group "nonspecific inflammatory arthritis" (NIA) consisted of cases with possible RA and others with a possible but unverified infectious background. Sera from 106 blood donors comprised the control material. Strains The reference strains used were MA PG6, MA strain Campo (previously M. hominis type 2) and M. fermentans G 11. Strain MA PG 6 was kindly supplied by Dr D. G.ff. Edward, Public Health Laboratory, Dulwich Hospital, London, and the other two by Dr Ruth Lemcke, Lister Institute, London. Strain Campo was originally isolated by Dr H. E. Morton. Strain 20-P was cultivated from synovial tissue and strain 176-M from joint fluid of definite RA patients, strain 338-M from bone marrow of a systemic lupus erythematosus (SLE) patient, and strain 71-M from a male with nongonococcal urethritis (NGU). Preparalion of antigens The antigens from the reference strains were prepared as described earlier ( l ) , but without phenol treatment. The isolates were grown in a medium containing pasteurized egg yolk (2).
(3). We decided to continue these investigations by
Indirect Hemagglutination ( I H A ) Technique This was performed as in earlier studies. The serum from each patient was studied simultaneously with different antigens and also before and after absorption with aggregated IgG. The first dilution made was always I : 16.
using several mycoplasma reference strains and hum a n isolates as antigens. It also seemed of interest t o discover whether absorption with aggregated IgG would affect the results.
Absorption uith aggregated 1gG The method described by other scientists ( 5 , 6) and commercial aggregated IgG from AG Behringwerke (Beriglobin) were used. Scand J Rheumatology 4
166
E . Junsson et al.
Table 1 . Correlation of mycoplasma IHA antibodies uith clinical disease MA=M. arthntidis PG 6 and Campo, MF=M. fermentans, 20-P and 176-M isolates from RA, 338-M from SLE, 71-T=a T-strain from NGU Number Total
Neg.
Pos.
Definite RA 2 yrs
18 12 19
10 4 9
8 8 10
6 2 4
1 3 4
6 6 9
3 2
49
23
26
12
8
21
6
5 3 3 11
3 3
2 3 3
2 2 2
3 -
1 3 1
-
8
6
3
5
1
23 25 12
11 10 5
12
7
9 8 1
4 4 4
8 11 6
4 4 -
Nonspecific inflammatory arthritis (1 yr 17 1-2 yrs 5 >2 yrs 12
6 3 6
11 2 6
11 2
4 1 I
10
2 5
5 2
15
19
13
6
17
7
Total
Scand J Rheumatol Downloaded from informahealthcare.com by Cornell University on 12/28/14 For personal use only.
Mycoplasma IHA antibodies in a titre of 3 1 6
Diagnosis and duration of the disease
Probable RA < I yr 1-2 yrs >2 yrs Total RA stage I RA stage I1 RA stage 111
Total
34
MF
MA
15
20, 176, 338
71
1
1 -
Osteoarthrosis Reiter's disease
antibodies. The series also included 2 patients with Reiter's disease and 4 with osteoarthrosis. One of the latter had antibodies against strain 20-P in a titre of 32. Twenty-one of the 26 patients in the definite RA group possessing mycoplasma antibodies showed titres of 16 or higher against isolates from RA and/or S L E , 12 against MA strain P G 6 and/or Campo, 8 against M. fermentans, and 6 against a n isolate from NGU (71-T). Antibodies against MA strain Campo were encountered more often than against MA strain PG6.
RESULTS Occurrence of mycoplasma IHA antibodies Table I reveals that out of 49 patients with definite RA, 26 had mycoplasma antibodies in titres of 16 or higher. Eight out of 12 subjects with adisease history of between one and two years possessed antibodies. There were 1 1 cases diagnosed as probable RA, 8 of them with positive findings. The third group consisted of 34 patients with possible RA or non-specific inflammatory arthritis (NIA). Nineteen of them showed mycoplasma
Table 2. Correlation of rnycoplasmri I H A antibody titres itith the duration of arthritic symptoms Number Total
Pos.
16-32
< I yr 1-2 yrs >2 yrs
40 '0 34
21 1; 19
10
Total
94
53
19
2 7
.
64-128
20-P 16-32
176-M 64-128
338-M a256
a256
16-32
64-128
a256
9 1
-
1
-
2 2 3
2 3 I
2 2 2
10 2 7
6 1 5
2 3 1
11
1
7
6
6
19
12
6
31 Scund J Rheumurology 4
M . fermentans
M . arfhriticlis
Duration of symptoms
I
19
37
Mycoplasma untibodies in K A
167
Table 3. Effect of absorption with aggregated I g G on rnycoplasma antibodies Total number of
Significant rise in titre
Diagnosis
rises
MA
Definite RA Probable RA Nonspecific inflam. arthritis
18 4 20
Total
42
decreases
8 3
MF
Significant decrease in titre
20, 176. 338
MA
MF
8 2
2 2
1 -
3 7
2
8
3
14
8 2
2 -
13
5
7
8
24
15
9
18
20, 176, 338
5 1
Scand J Rheumatol Downloaded from informahealthcare.com by Cornell University on 12/28/14 For personal use only.
Table 4. Details of mycoplusmu antibody titres in some patients Negative=titres under 16. First line=antibodies before absorption with aggregated IgG, 2nd line=antibodies after absorption with aggregated IgG
No. Diagnosis
Duration of disease
Stage
Latex
WaalerRose
39
Definite RA
>2 yrs
I1
-
2 yrs
111
++
1 000
43
Definite R A
1-2 yrs
I1
-
Mycoplasma antibodies
2 yrs
I
35
Probable RA
1-2 yrs
I
++ +++ ++
29
Prohable RA
1-2 yrs
I
-
MYCOPLASMA ANTIBODIES IN RHEUMATOID ARTHRITIS Elli Jansson, Paavo Makisara and Sirkka Tuuri
Scand J Rheumatol Downloaded from informahealthcare.com by Cornell University on 12/28/14 For personal use only.
From the University of Tampere, School of Medicine, 33520 Tampere, and Rheumatism Foundation Hospital, 18120 Heinola, Finland
ABSTRACT. Sera of 100 patients under examination at the Outpatient Department of the Rheumatism Foundation Hospital were studied by the indirect hemagglutination technique, using both mycoplasma reference strains, and isolates from RA and SLE as antigens. The series consisted of five groups: I, definite RA (49 patients); 11, probable RA (11);111, possible RA or nonspecific inflammatory arthritis (34); IV, osteoarthrosis (2); V, Reiter's disease (4). Mycoplasma antibodies in titres of 16 or higher were encountered in groups I-IV in 26,8,19, and one case respectively. Twentyone out of 106 blood donors had antibodies against an isolate from RA and/or M. urthn'tidis strain PG 6. The titres found were 16 or 32, except in two cases, 128. In the definite RA group, 21 out of 26 patients possessing mycoplasma antibodies, showed titres of 16 or higher against isolates from RA and/or SLE, 12 against M. urthritidis strain PG 6 and/or Campo, 8 against M. fermentuns, and 6 against a T-strain from NGU. Antibodies against M. arthn'tidis strain Campo were found more often than against strain PG 6. The longer the duration of the arthritic symptoms was, the more frequent seemed also to be the occurrence of mycoplasma antibodies.
Antibodies against Mycoplasma arthritidis (MA) strain Campo, M. fermentans and a f e w isolates from human rheumatoid arthritis (RA) were investigated earlier in sera from patients with this disease. Complement-fixing or metabolism-inhibiting antibodies were not usually detected (2). However, low antibody titres against a human isolate (strain 176-M) were observed by the indirect hemagglutination (IHA) technique. Out of 421 sera with a Waaler-Rose titre of 2128, 30% possessed antibodies in a titre of 2 8 , a n d 19% in a titre of 216
M A T E R I A L AND METHODS Sera These were collected from patients under examination at the Outpatient Department of the Rheumatism Foundation Hospital, Heinola. The series is not entirely unselected as the proportion of relatively new and obscure cases is accented. Patients fulfilling at least five ARA criteria were classified as definite RA, those with a minimum of three as probable RA. The RA group was also subdivided by ARA criteria concerning anatomical joint changes (7). The group "nonspecific inflammatory arthritis" (NIA) consisted of cases with possible RA and others with a possible but unverified infectious background. Sera from 106 blood donors comprised the control material. Strains The reference strains used were MA PG6, MA strain Campo (previously M. hominis type 2) and M. fermentans G 11. Strain MA PG 6 was kindly supplied by Dr D. G.ff. Edward, Public Health Laboratory, Dulwich Hospital, London, and the other two by Dr Ruth Lemcke, Lister Institute, London. Strain Campo was originally isolated by Dr H. E. Morton. Strain 20-P was cultivated from synovial tissue and strain 176-M from joint fluid of definite RA patients, strain 338-M from bone marrow of a systemic lupus erythematosus (SLE) patient, and strain 71-M from a male with nongonococcal urethritis (NGU). Preparalion of antigens The antigens from the reference strains were prepared as described earlier ( l ) , but without phenol treatment. The isolates were grown in a medium containing pasteurized egg yolk (2).
(3). We decided to continue these investigations by
Indirect Hemagglutination ( I H A ) Technique This was performed as in earlier studies. The serum from each patient was studied simultaneously with different antigens and also before and after absorption with aggregated IgG. The first dilution made was always I : 16.
using several mycoplasma reference strains and hum a n isolates as antigens. It also seemed of interest t o discover whether absorption with aggregated IgG would affect the results.
Absorption uith aggregated 1gG The method described by other scientists ( 5 , 6) and commercial aggregated IgG from AG Behringwerke (Beriglobin) were used. Scand J Rheumatology 4
166
E . Junsson et al.
Table 1 . Correlation of mycoplasma IHA antibodies uith clinical disease MA=M. arthntidis PG 6 and Campo, MF=M. fermentans, 20-P and 176-M isolates from RA, 338-M from SLE, 71-T=a T-strain from NGU Number Total
Neg.
Pos.
Definite RA 2 yrs
18 12 19
10 4 9
8 8 10
6 2 4
1 3 4
6 6 9
3 2
49
23
26
12
8
21
6
5 3 3 11
3 3
2 3 3
2 2 2
3 -
1 3 1
-
8
6
3
5
1
23 25 12
11 10 5
12
7
9 8 1
4 4 4
8 11 6
4 4 -
Nonspecific inflammatory arthritis (1 yr 17 1-2 yrs 5 >2 yrs 12
6 3 6
11 2 6
11 2
4 1 I
10
2 5
5 2
15
19
13
6
17
7
Total
Scand J Rheumatol Downloaded from informahealthcare.com by Cornell University on 12/28/14 For personal use only.
Mycoplasma IHA antibodies in a titre of 3 1 6
Diagnosis and duration of the disease
Probable RA < I yr 1-2 yrs >2 yrs Total RA stage I RA stage I1 RA stage 111
Total
34
MF
MA
15
20, 176, 338
71
1
1 -
Osteoarthrosis Reiter's disease
antibodies. The series also included 2 patients with Reiter's disease and 4 with osteoarthrosis. One of the latter had antibodies against strain 20-P in a titre of 32. Twenty-one of the 26 patients in the definite RA group possessing mycoplasma antibodies showed titres of 16 or higher against isolates from RA and/or S L E , 12 against MA strain P G 6 and/or Campo, 8 against M. fermentans, and 6 against a n isolate from NGU (71-T). Antibodies against MA strain Campo were encountered more often than against MA strain PG6.
RESULTS Occurrence of mycoplasma IHA antibodies Table I reveals that out of 49 patients with definite RA, 26 had mycoplasma antibodies in titres of 16 or higher. Eight out of 12 subjects with adisease history of between one and two years possessed antibodies. There were 1 1 cases diagnosed as probable RA, 8 of them with positive findings. The third group consisted of 34 patients with possible RA or non-specific inflammatory arthritis (NIA). Nineteen of them showed mycoplasma
Table 2. Correlation of rnycoplasmri I H A antibody titres itith the duration of arthritic symptoms Number Total
Pos.
16-32
< I yr 1-2 yrs >2 yrs
40 '0 34
21 1; 19
10
Total
94
53
19
2 7
.
64-128
20-P 16-32
176-M 64-128
338-M a256
a256
16-32
64-128
a256
9 1
-
1
-
2 2 3
2 3 I
2 2 2
10 2 7
6 1 5
2 3 1
11
1
7
6
6
19
12
6
31 Scund J Rheumurology 4
M . fermentans
M . arfhriticlis
Duration of symptoms
I
19
37
Mycoplasma untibodies in K A
167
Table 3. Effect of absorption with aggregated I g G on rnycoplasma antibodies Total number of
Significant rise in titre
Diagnosis
rises
MA
Definite RA Probable RA Nonspecific inflam. arthritis
18 4 20
Total
42
decreases
8 3
MF
Significant decrease in titre
20, 176. 338
MA
MF
8 2
2 2
1 -
3 7
2
8
3
14
8 2
2 -
13
5
7
8
24
15
9
18
20, 176, 338
5 1
Scand J Rheumatol Downloaded from informahealthcare.com by Cornell University on 12/28/14 For personal use only.
Table 4. Details of mycoplusmu antibody titres in some patients Negative=titres under 16. First line=antibodies before absorption with aggregated IgG, 2nd line=antibodies after absorption with aggregated IgG
No. Diagnosis
Duration of disease
Stage
Latex
WaalerRose
39
Definite RA
>2 yrs
I1
-
2 yrs
111
++
1 000
43
Definite R A
1-2 yrs
I1
-
Mycoplasma antibodies
2 yrs
I
35
Probable RA
1-2 yrs
I
++ +++ ++
29
Prohable RA
1-2 yrs
I
-
