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Year : 2013  |  Volume : 31  |  Issue : 2  |  Page : 177--179

Anti-ganglioside antibodies in patients with Guillain Barré syndrome and other neurological disorders

C Vaishnavi1, C Behura1, S Prabhakar2, A Sharma2, P Kharbanda2,  
1 Department of Gastroenterology, Post Graduate Institute of Medical Education and Research, Chandigarh, India
2 Department of Neurology, Post Graduate Institute of Medical Education and Research, Chandigarh, India

Correspondence Address:
C Vaishnavi
Department of Gastroenterology, Post Graduate Institute of Medical Education and Research, Chandigarh
India

Abstract

A study was performed on 59 Guillain-Barré syndrome (GBS) cases, 58 neurological controls (NC) and 60 non-neurological controls (NNC) to investigate the association of anti-ganglioside antibodies in GBS and other neurological disorders. Campylobacter jejuni was isolated from 5.7% of GBS patients. Anti-ganglioside immunoglobulin G was present in 82% and immunoglobulin M in 46% in acute inflammatory demyelinating polyneuropathy patients, 70% and 44% respectively in acute motor axonal neuropathy subgroup and 38% each in acute motor sensory axonal neuropathy subgroup. Though high intensity of anti-gangliosides was present in the GBS patients, the NC patients also had adequate anti-gangliosides compared with the NNC group.

How to cite this article:
Vaishnavi C, Behura C, Prabhakar S, Sharma A, Kharbanda P. Anti-ganglioside antibodies in patients with Guillain Barré syndrome and other neurological disorders.Indian J Med Microbiol 2013;31:177-179

How to cite this URL:
Vaishnavi C, Behura C, Prabhakar S, Sharma A, Kharbanda P. Anti-ganglioside antibodies in patients with Guillain Barré syndrome and other neurological disorders. Indian J Med Microbiol [serial online] 2013 [cited 2020 Oct 26 ];31:177-179
Available from: https://www.ijmm.org/text.asp?2013/31/2/177/115223

Full Text

 Introduction



Guillain-Barré syndrome (GBS) is an autoimmune, polyradiculoneuropathy affecting the peripheral nervous system, usually triggered by an acute infectious agent in 50-70% of cases. The targets of such immune attack are thought to be gangliosides present in large quantities on human nerve tissues. Recent studies suggest that Campylobacter jejuni may be the most common preceding pathogen for GBS. [1] Kaldor and Speed, [2] found that 38% of GBS patients had recent C. jejuni infection. GBS is generally precipitated 1-3 weeks following C. jejuni infection suggesting a humoral immunopathogenic mechanism.

Autoantibodies against gangliosides are found in patients with peripheral neuropathies, including GBS. In GBS, both immunoglobulin M (IgM) and immunoglobulin G (IgG) against GM1, GD1a and GT1b and only IgG antibodies against GQ1b are found in Miller-Fisher syndrome. [3] In case of multi-focal motor neuropathy, IgM antibodies against GM1 ganglioside are found [4] and with sensory neuropathy IgG antibodies against GD1b are found. [5] In the present study, patients with GBS were investigated for C. jejuni infection and anti-ganglioside antibodies. Control subjects comprised of two groups of patients - one with other neurological ailments and the second with non-neurological conditions.

 Patients and Methods



Patients attending the Neurology Out-patient Department or other wards of a tertiary care teaching hospital were enrolled for prospective investigation between November 2006 and October 2009. The study was approved by the Institute Ethical Committee and informed consent was taken from all patients.

The study population was divided into three groups: (i) GBS patients (n = 59) who underwent nerve conduction studies to establish the diagnosis [6] and electrophysiological criteria, [7] to classify the GBS patients (ii) non-paralytic neurological patients (n = 58) with symptoms such as back pain, migraine, dizziness, muscle weakness, multiple sclerosis, trembling, loss of memory, brain tumour, etc., served as neurological controls (NC) to the GBS group (iii) non-neurological patients (n = 60) acted as non-neurological controls (NNC) to GBS and NC groups.

Faecal samples obtained from 35 GBS, 41 NC and 42 NNC patients were cultured directly and by filtration method on selective campylobacter blood agar. Colonies grown were identified by standard methods inclusive of hippurate hydrolysis. Serogrouping of culture isolates was performed by Penner's method using C. jejuni antisera (Denka Seiken, Japan).

Sera collected from patients were tested for the presence of IgG and IgM antibodies to gangliosides using commercially available EUROLINE test system (Lubeck, Germany). Briefly, each serum diluted 1:51 in buffer was incubated with membrane test strips printed with purified ganglioside fraction (GM1, GM2, GM3, GD1a, GD1b, GT1b and GQ1b) and rocked for 1 h for binding to their specific antigens. After washing with buffer solution, enzyme conjugate was added and again rocked for 1 h. Wash cycles with buffer solution were repeated 3 times and substrate solution was added to it. After rocking for 10 min, the strips were washed and fixed on an evaluation protocol to read the colour reaction with the enzyme-coupled secondary antibody as per the manufacturer's protocol. The Fisher's exact test was carried out to compare between the IgG and IgM in all three groups as well as between each group.

 Results



Based on clinical examination and electrophysiological findings, the GBS group comprised of 28 patients with acute inflammatory demyelinating polyneuropathy (AIDP), 23 with acute motor axonal neuropathy (AMAN) and 8 with acute motor sensory axonal neuropathy (AMSAN).

Of 35 patients, Campylobacter Penner group O:19 was isolated from the stool samples of two (5.7%) GBS patients; but, none from either of the NC and NNC groups.

Based on signal intensity, the immunoblot results were divided into negative (N) when there was no signal, borderline (B) with very weak band, positive (P) with medium to strong band and strongly positive (SP) when the band was very strong with an intensity comparable to the control band. [Figure 1] shows the percentage distribution of anti-ganglioside antibodies in the different groups. There was no significant difference in the anti-ganglioside antibodies between the GBS and the NC group. Very weak bands of GT1b were detected in 6.7% of NNC patients. [Figure 2] shows the distribution of IgG and IgM anti-ganglioside antibodies in the GBS subgroups. IgG antibodies against gangliosides were present in 23/28 (82%) and IgM antibodies in 13/28 (46%) in AIDP patients. In the AMAN subgroup, IgG was present in 16/23 (70%) and IgM in 10/23 (44%). In AMSAN subgroup, both antibodies were present in 3/8 (38%) of the samples. [Table 1] shows the statistical comparison of IgG and IgM between the different groups of patients investigated.{Table 1}{Figure 1}{Figure 2}

 Discussion



Infection with C. jejuni is a common antecedent to GBS and might play a role in initiating demyelination resulting in neuropathies. Serological studies have shown the occurrence of C. jejuni enteritis in 13% to 72% of GBS patients. [8],[9] In the present study, both IgM and IgG antibodies against GM1, GM2 and GM3 ranging from 9% to 24% in GBS and NC groups, but none in the NNC group were detected. It has been reported that GM1 antibodies, mainly IgM, are associated with multi-focal motor neuropathy with prevalence of 40% to 70%. [4] Elevated antibody titres against GM1 also occur in 22% to 30% GBS cases and the titre increases to a maximum value in the acute phase and decreases as the disease progresses.

It is known that IgG anti-GD1a antibody is a predictor of the motor axonal form of GBS. Dikmen and Oge, [10] reported recurrent GBS with GD1a anti-ganglioside antibodies in a woman with an episode of gastroenteritis. GD1b antibodies occur in rare cases of patients with sensory neuropathy, whereas GQ1b is detected in >90% of patients with Miller-Fisher syndrome. [11] GM1, GD1a and GT1b are known to be associated with GBS. [11] A very high association of these three gangliosides was found in our GBS patients. Antibodies against GD1b were found in 2% to 17% in GBS and NC groups, but none in the NNC group. Only GT1b was detected in 6.7% of NNC patients as very weak bands.

Kuroki et al.,[12] reported the frequency of PEN 19 strains isolated from patients with GBS. Even in the present study, the C. jejuni isolated were of Penner O:19 type. The mean excretion time of C. jejuni in stools is only 16 days and patients generally report much later after the onset of neuropathy, resulting in lower isolation rate. Lipo-oligosaccharide structures on C. jejuni surface can elicit an autoimmune-mediated attack against host peripheral neural tissue through molecular mimicry.

Antibodies to C. jejuni may remain elevated for several weeks after acute infection. Both IgM and IgG antibodies against GM1, GM2 and GM3 were detected in GBS and NC groups, but none in the NNC group. GM1, GD1a and GT1b, known to be associated with GBS were found in our GBS patients. However, NC patients also had a high level of several of these anti-ganglioside antibodies. Triggering agents for anti-ganglioside antibodies are C. jejuni and celiac diseases. [13] Celiac disease was not ruled out in our patients, which could have been one of the factors for the presence of anti-ganglioside antibodies.

 Conclusion



In conclusion, though high intensity of anti-gangliosides were present in the GBS patients, the NC patients also had adequate anti-ganglioside antibodies compared with the NNC group, which could be triggered by factors other than C. jejuni.

 Acknowledgment



The authors (CV, SP and PK) are grateful to the Indian Council of Medical Research, New Delhi, for the funding. Mr. Baljit Singh is acknowledged for technical help and Dr. P.K. Gupta for the statistical evaluation.

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