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ORIGINAL ARTICLE
Year : 2005  |  Volume : 22  |  Issue : 3  |  Page : 77-80 Table of Contents   

Specific IgG and its subclass antibodies after immunotherapy with gynandropsis gynandra


Institute Immunology Unit, Bhagawan Mahavir Medical Research Centre, 10-1-1, Mahavir Marg Hyderabad-500 004., India

Correspondence Address:
K.J.R Murthy
Institute Immunology Unit, Bhagawan Mahavir Medical Research Centre, 10-1-1, Mahavir Marg Hyderabad-500 004.
India
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Source of Support: None, Conflict of Interest: None


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   Abstract 

Background : About 10 to 15 % of the Indian population is known to suffer from major allergic disorders such as Asthma, Rhinitis, Atopic Dermatitis and Urticaria. Aeroallergens play a major role in the pathogenesis of respiratory allergic diseases. Among the aeroallergens, pollens are major causative agents. The predominance of pollen allergens necessitate the need to assess the specific immunotherapy (SIT) in allergic patients.
Objective : To evaluate the effect of immunotherapy based on the presence of IgG and its subclass antibodies towards whole pollen antigen of Gynandropsis gynandra (G.gynandra) and its fractions.
Material and Methods : A study was conducted in 30 bronchial asthma patients on immunotherapy, by assessing the levels of IgG and its subclasses specific to G. gynandra pollen.
Results : There was a significant increase in IgG and its subclass antibodies to whole pollen antigen and its fractions i.e. >90kD, 46­37kD and 36-32kD after the course of IT.
Conclusion : The use of peptide fractions may be more appropriate instead of the whole pollen antigen to test the effect of immunotherapy.

Keywords: Immunotherapy (IT), Gynandropsis gynandra, IgG, IgG1, IgG2, IgG3 and IgG4. Lung India 2005; 22: 77-80


How to cite this article:
Latha G S, Lakshmi V V, Rani H S, Anuradha B, Murthy K. Specific IgG and its subclass antibodies after immunotherapy with gynandropsis gynandra. Lung India 2005;22:77-80

How to cite this URL:
Latha G S, Lakshmi V V, Rani H S, Anuradha B, Murthy K. Specific IgG and its subclass antibodies after immunotherapy with gynandropsis gynandra. Lung India [serial online] 2005 [cited 2019 Jun 17];22:77-80. Available from: http://www.lungindia.com/text.asp?2005/22/3/77/44448


   Introduction Top


About 10 to 15 % of the Indian population is known to suffer from major allergic disorders such as Asthma, Rhinitis, Atopic Dermatitis and Urticaria. [1] There are many reports on predominance of different pollens in different parts of India. Shivpuri and Prakash observed Prosopis juliflora as a major pollen allergen based on the skin test positive reaction in 12% of the patients in Delhi, [2] however geographical variations are known to occur. Santosh et al reported Gynandropsis gynandra, Parthenium hysterophorous, Prosopis juliflora, Pennisetum as the most allergenic pollens in patients residing in and around Hyderabad city. [3] The predominance of pollen allergens necessitate the need to assess the specific immunotherapy (SIT) in allergic patients.

Pollen allergy is mainly caused by proteins, glycoproteins or even single peptide present in the pollen wall and cytoplasm. Currently whole pollen allergen extracts are used for immunotherapy. There is a need to identify and isolate the appropriate major allergenic fractions, which could be used for SIT. The present study is designed to evaluate the effect of immunotherapy based on the presence of IgG and its subclass antibodies to whole pollen antigen and fractions of Gynandropsis gynandra (G.gynandra).


   Material and Methods Top


Subjects and Clinical studies

Thirty patients diagnosed as bronchial asthma attending the Allergy Clinic at Mahavir Hospital & Research Centre were studied. The criteria used to determine the atopy in these patients were bronchial asthma with elevated total sIgE levels and positive skin test responses. Patients below 12 years and above 60 years and pregnant women were excluded. Informed consent was taken prior to skin test and blood collection. Ethical permission was obtained from the Institutional Ethics Committee to carry out the study.

Immunotherapy

Patients with positive skin tests were put on an immunotherapy for a period of 18 months. Patients who underwent SIT with G. gynandra were included in the study. The study did not exclude patients who received SIT for other allergens in addition to G. gynandra. SIT consisted of subcutaneous injections of antigen(s), by gradually increasing the dose of antigen from 1:5000 through 1:500 to 1:50 covered over a period of 18 months (Cure well India Ltd).

Pollen : The proteins were extracted from the pollens of G.gynandra as described by Sridhara et al. [4]

Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE): SDS-PAGE was performed with the extracted proteins, using a vertical slab gel apparatus (Bio-rad, laboratories) as described by Laemmli. [5] Ten fractions were isolated from the gel by Electro elution as described by Andersen P. [6] The fractions with different mol.wts were fraction 1 (>90kD), fraction 2 (89-70kD), fraction 3 (69-65kD), fraction 4 (64-56kD), fraction 5 (55-47kD), fraction 6 (46-37kD), fraction 7 (36­32kD), fraction 8 (31-22kD), fraction 9 (21-14kD) and fraction 10 (<14kD).

Specific IgG and its subclass antibodies: ELISA was performed to measure the G.gynandra­ specific IgG and its subclass antibody levels in individual patients sera before and after the course of immunotherapy by the method of Voller et al. [7]

Statistical Analysis

Specific IgG and its subclass antibody levels with whole antigen and its fractions were analysed after the course of immunotherapy by student's 't' test.


   Results Top


The IgG and subclasses IgG1, IgG2, IgG 3 and IgG 4 antibodies were significantly high to the whole antigen and its fractions 1, 6 and 7 after the course of IT. [Table 1],[Table 2],[Table 3],[Table 4],[Table 5] respectively.


   Discussion Top


The exact immunomodulatory mechanism by which immunotherapy switches off allergies is uncertain. It has been postulated that initially specific IgG 1 antibodies are provoked and later specific IgG 4 antibodies are induced towards the offending allergen. An associated reduction in mucosal mast cell numbers and a decrease in antigen-induced eosinophil migration to the site of inflammation are noted during immunotherapy and modulate the T-helper cells, causing switching from predominantly Th2 (IgE inducing) to Th1 (IgG inducing) subsets and as a result of this allergen-specific IgE falls with successful immunotherapy. [8] In the present study with whole pollen antigen, IgG antibodies were increased after the course of IT. Similar observation was also reported by Birkner et al., for birch pollen allergic patients. [9] The increase in humoral responses could be because of the development of antigen-specific immunoglobulin (IgG), including subclasses which may be associated with specific responses in improving airways and allergic responses. However, changes in humoral immune function during immunotherapy are not associated temporally with improvement in clinical responses, and the actual role of such potential "blocking" antigen-specific IgG molecules remains under skepticism. [10]

The subclass blocking antibodies i.e IgG1, IgG2,IgG3 and IgG4 were increased after the course of IT in the study. Park et al., reported an increase in IgG 1 and IgG 4 antibodies after the course with Hop Japanese pollen. [11] Gehlhar et al has reported in grass pollen IT group an increase in IgG 1 , IgG 2 and IgG 4 antibodies after the course. [12] Fling et al., have reported an increase in IgG, IgG 1 , IgG 4 and no change in IgG 2 and IgG 3 after the course with mountain cedar pollen. [13] While Brunet et al., demonstrated that the blocking antibody responses were restricted to only IgG 1 and IgG 4 after the course in rye grass pollen IT patients. [14] But in the present study the IgG 1 , IgG 2 , IgG 3 and IgG 4 antibody responses were found after the course of IT.

There are few studies demonstrating clinical improvement in the absence of an increase in allergen­specific IgG, but the bulk of evidence shows an inverse correlation between reduced symptom, medication scores and serum levels of specific IgG. Moss et al., have reported that a competition exists between IgE and IgG antibody responses to distinct antigens in individuals and called this as "Parallel antigen" recognition. [15] There has been a lack of knowledge on the control mechanism and kinetics of allergen-specific IgG subclass antibodies. Though the changes in B cell and T cell responses, especially IgG antibodies and T (H) 1/T (H) 2 cytokine production, may be the major mechanism underlying the clinical efficacy of allergen immunotherapy and in the prevention of the development of allergic phenotypic changes and multiple mechanisms may be involved in the outcome of alterations of the natural course of allergic disorders.

The results of the present study demonstrate that with whole pollen antigen, the IgG and subclass antibodies were found to be significantly increased and also fractions with mol.wt >90kD, 46-37kD, 36-32kD have shown a highly significant increase after the course. So instead of studying the whole pollen antigen, it would be more appropriate to identify atopy by using peptide fractions or epitopes rather than general antigen in identifying atopy as well as for effect of immunotherapy.


   Acknowledgements Top


We are grateful to Lady Tata Memorial Trust & Council of Scientific and Industrial Research (CSIR) for the financial support during the study. We thank Dr.Akbar Yazdani, Dr. Nafees Sultana & Ms. R. Eswari for the clinical and technical help rendered during the study.

 
   References Top

1.Vishwanathan R, Definition, incidence, etiology and natural history of asthma. Ind J Chest Dis 1964; 6: 108-124.  Back to cited text no. 1    
2.Shivpuri DN, and D Parkash. A study in allergy to Prosopis juliflora (Kabuli Keekar). Ann Allergy 25: 643-648, 1967.  Back to cited text no. 2    
3.Santosh Reddy P, Aerobilogical survey of Hyderabad. Thesis presented to the Osmania University for the award of Ph.D in Botany 1990.  Back to cited text no. 3    
4.Sridhara S, BP Singh, Naveen Arora, Jyotsna Verma and SV Gangal. A study of antigenic and allergenic changes during storage in three different biological extracts. Asian Pacific J Allergy Immunol. 1992; 10: 33-38.  Back to cited text no. 4    
5.Laemmli UK. Cleavage of structural proteins during assembly of the head of the bacteriophage T4. Nature 1970; 227:680-685.  Back to cited text no. 5  [PUBMED]  
6.Andersen P, and I Heron Simultaneous electroelution of whole SDS-polyacrylamide gels for the direct cellular analysis of complex protein mixtures. J Immunol 1993;161: 29-39.  Back to cited text no. 6    
7.Voller A, Bidwell D, Barlett A Enzyme linked immunosorbent assay; In; Rose NR, Friedman H, (eds): Manual of Clinical Immunology. Washington, American Society for Microbiology, 1980.  Back to cited text no. 7    
8.Akdis CA, Blaser K.Mechanisms of allergen-specific immnotherapy. Allergy 2000; 55:522--530.  Back to cited text no. 8  [PUBMED]  [FULLTEXT]
9.Birkner T, Rumpold H, Jarolim E, Ebner H, Breitenbach M, Skvaril F, Scheiner O, Kraft D. Evaluation of immunotherapy- induced changes in specific IgE, IgG and IgG subclasses in birch pollen allergic patients by means of immunoblotting. Correlation with clinical response. Allergy 1990 ; 45: 418-26.  Back to cited text no. 9    
10.Robinson D, Q Hamid, S Ying, A Tsicopoulos, J Barkans, A M Bentley, C Corrigan, S R Durham, and A B Kay. Predominant Th2-like bronchoalveolar T-lymphocyte population in atopic asthma. N Engl J Med 1992; 326: 298-304.  Back to cited text no. 10    
11.Park HS, Nahm DH, Kim HY, Suh YJ, Cho JW, Kim SS, Lee SK, Jung KS. Clinical and immunologic changes after allergen immunotherapy with Hop Japanese pollen. Ann Allergy Asthma Immunol 2001; 86: 444-8.  Back to cited text no. 11  [PUBMED]  
12.Gehlhar K, Schlaak M Becker, W Bufe A. Monitoring allergen immunotherapy of pollen-allergic patients: the ratio of allergen-specific IgG4 to IgG1 correlates with clinical outcome. Clinical&Experimental Allergy. 1999;29: 497-506.  Back to cited text no. 12    
13.Fling JA, Ruff ME,Parker WA. Suppression of the late phase cutaneous response by immunotherapy. J Allergy Clin Immunol 1989;83: 101-109.  Back to cited text no. 13    
14.Brunet C, Berdard PM, Lavoie A, Jobin M, Hebert J. Allergic rhinitis to ragweed pollen. I. Reassessment of the effects of immunotherapy on cellular and humoral responses. J Allergy Clinic Immunol 1992 ; 89 (1Pt 1): 76-86.  Back to cited text no. 14    
15.Moss RB, Hsu YP, Kwasnicki JM, et al Isotypic and antigenic restriction of the blocking antibody response to ryegrass pollen: Correlation of rye group I antigen-specific IgG1 with clinical response. J Allergy Clin Immunol 1987; 79: 387-398.  Back to cited text no. 15    



 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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    Introduction
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