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Year : 1990  |  Volume : 56  |  Issue : 2  |  Page : 133-135

Phagocytic activity and the response to autovaccines in recurrent staphylococcal boils

Correspondence Address:
A C Karnik

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The present study was aimed to see the relationship between the phagocytic function as determined by NBT reduction test and the response to the staphylococal boils. Of the 34 cases of recurrent staphylococcal cutaneous infections, 19 showed normal phagocytic activity while 2 had marginally diminished and 13 had grossly diminished phagocytic activity. Sixteen of the vaccines (84%) with normal phagocytic funetion responded well to the vaccine therapy while only 3 of the 13 (23%) with poor phagocytic function showed the beneficial effects of the autovaccines. Thus the findings recommend the use of staphyloccoccal autovaccines in the cases with adequate phagocytic activity as seen by NBT reduction test but not as much for those with inadequate phagocytic function.

Keywords: Staphylococcal infections, Phagocytic activity, Autovaccine therapy

How to cite this article:
Karnik A C, Vijayvargiya R, Chitnis D S, Sethi N C. Phagocytic activity and the response to autovaccines in recurrent staphylococcal boils. Indian J Dermatol Venereol Leprol 1990;56:133-5

How to cite this URL:
Karnik A C, Vijayvargiya R, Chitnis D S, Sethi N C. Phagocytic activity and the response to autovaccines in recurrent staphylococcal boils. Indian J Dermatol Venereol Leprol [serial online] 1990 [cited 2021 Jan 15];56:133-5. Available from:

The use of autogenous vaccines against recurrent staphylococcal boils is as old as the century.[1],[2],[3] However, these vaccines got a setback in the past few years because of some controversial results. In our own experience a significant proportion of the patients bene­fitted from the vaccines whereas some continued to have recurrence of the staphylococcal boils. It was thought that the failure of the vaccine was due to inadequate phagocytic functions. The present study was carried out to verify it.

  Materials and Methods Top

Thirty four patients with recurrent staphylo­coccal boils were included in the study. The patients had been having recurrent boils for more than two months, and had not shown good response to antimicrobials selected on the basis of in vitro drug susceptibility. The patients were between 6-64 years of age. Out of these 34 patients, 30 had recurrent boils and 4 had recurrent pyoderma. Phagocytic function was assessed by the nitroblue tetrazolium (NBT) dye reduction test.[4]

  NBT dye reduction test Top

To 5 ml heparinized blood, 2.5 ml of 6% dextran, MW 70,000 (Ralley's India) was added and kept at 37°C for 30 minutes. Plasma, rich in leukocytes was separated and centrifuged at 2000 rpm for 5 minutes. The cell pellet contain­ing the granulocytes was washed with Hank's balanced salt solution (HBSS) from Hi Media Lab (Bombay) 2-3 times and then suspended in 0.4 ml Candida cells (10 6sub cells), 0.2 nil of 0.2% NBT (Sigma, USA) solution and 0.1 ml of autologous plasma. The control contained HBSS in place of Candida cells. Both the control and the test were kept at 37°C for one hour and then visualized under the microscope for the blue coloured Formazon in the granulocytes. NBT reduction observed in 40% or less of the activated neutrophils was considered as inade­quate phagocytic activity, while between 40-60% was interpreted as marginally diminished phago­cytosis. Reduction of NBT by more than 60% of the neutrophils was reported as normal phagocytosis.

  Autovaccine preparation Top

This was based on the protocol used at Haffkine's Institute, Bombay, using Staphylo­coccus aureus isolated from the pus of the respec­tive patient. Growth on nutrient agar was harvested in 0.25% carbol saline and the bacterial density was adjusted to 10 8sub cells/ml. The bacterial cells were killed by exposure to heat at 70-75°C for 90 minutes. The heated suspen­sion was tested for sterility. The intradermal test was performed in every case. The vacci­nation schedule consisted of gradually increasing doses of the autovaccine from 0.1 ml to 1 ml and were given as deep intramuscular injections at 3-4 day intervals. After 10 such injections, booster doses of 1 ml were administered at weekly intervals for 3-6 weeks.

  Results Top

Response of the vaccines to the autovaccine is depicted in [Table - 1]. Sixteen (84%) of the 19 cases with normal phagocytic activity showed disappearance of the boils, and there was no recurrence during the one year of follow up. Even the two cases with marginally diminished phagocytic activity responded well to the auto­vaccine therapy. However, in cases with grossly reduced phagocytic function the bene­ficial effect was seen in only 3 (23%) cases. Thus, the phagocytic function when assessed with NBT dye reduction test, correlated well with the response to the staphylococcal auto­vaccine.

  Comments Top

Staphylococcus aureus elaborates an array of enzymes such as lipases, esterases, hyaluro­nidase and toxins like alpha, beta and deltas which are antigenic. However, antibody res­ponses to these antigens were not studied. Thus, the quantum of antibodies to them in recurrent infections is not known. Similarly, the anti­body levels after the hyperimmunization effect of the vaccines were not determined. This would be the next phase of work.

The main events in the defence from pyogenic infections are : chemotaxis which attracts the phagocytes at the site of infection,[7],[8] opsoni­sation where the organisms get coated with the antibody and become susceptible for engulfment by the phagocytes[9],[10],[11] and intracellular killing due to the burst of lytic enzymes from phago­somes.[12],[13] The chemotactic defect, was not the likely cause in our cases, because in most of them huge boils with abundance of pus were present. In some of the cases agglutinating antibodies to Staphylococcus were checked and found to be present in a titre above 1 : 40. This suggests that antibodies to the surface antigens of staphylococci which can act as opsonins are often present even before the autovaccine therapy and hence the lack of opsonizing antibodies does not seem to be the cause of the recurrent infection.

Refractoryness to the intracellular killing enzymes could be an important factor for the recurrence of staphylococcal infections.[14] Weaker peroxidase burst in the phagocytes is also likely to be responsible for the survival of staphylococcal cells inside the phagocytes.[14] Intracellular killing of the staphylococci within the phagocytes was not determined in our study, however, the NBT reduction test shows the activity of peroxidases and has been shown to­correlate with intracellular killing capacity.[15],[16],[17] The defect in the reduction of NBT dye by the phagocytes correlated very well with the response to the autovaccines in the present work. In other words, the normal phagocytic function was required to have the benefits of the autovaccine. The mechanism of action of the autovaccine remains highly speculative, it could be due to the antibodies directed against the toxins or the damaging enzymes. Hypersensitivity to staphy­lococcal antigen is considered to be the mecha­nism of pathogenesis.[18],[19] The autovaccitie therapy being a hyperimmunization process, would generate excess of specific IgG antibodies that can mask the action of IgE antibodies. Evidence for these speculations however, were not studied in the present study.

  References Top

1.Wright AE and Douglas SR : An experimental investigation of the role of the body fluids in connection with phagocytosis, Proc Roy Soc Lond, 1903; 72 : 357.  Back to cited text no. 1    
2.McCoy KL and Kennedy ER : Autogenous vaccine therapy in staphylococcal infections, JAMA, 1960; 74 : 35-38.  Back to cited text no. 2    
3.Neter E, Rajnovich E and Goznski EA : Study of staphylococcal antibodies of Rantz type placen­tal transfer and titres in sera of children of various ages, Pediat, 1960; 25 : 21-26.  Back to cited text no. 3    
4.Segal DW : Nitroblue tetrazolium test, Lancet, 1974; ii : 1245-1252.  Back to cited text no. 4    
5.Brown JJ : J Pathol Bacteriol, 1960; 79 : 257, Quoted by Smith DT, Conant NF and Overman JR : The micrococci, in: Microbiology, Thirteenth ed, Editor, Zinsser 14 : Meredith Publishing Company, USA, 1964; p 436-499.  Back to cited text no. 5    
6.Glenndy Al and Stevens MF : J Pathol and Bacteriol, 1952 ; 66 : 187, Quoted by Smith DT, Conant NF and Overman JR :The micrococci, in : Microbiology, Thirteenth ed, Editor, Zinsser, H : Meredith Publishing Company, USA, 1964: p 436-499.  Back to cited text no. 6    
7.Zimond SH : Chemotaxis of polymorphonuclear leukocytes, J Cell Biol, 1978; 77 : 269-271.  Back to cited text no. 7    
8.Keller HU and Sorkin E : Chemotaxis of leuko­cytes, Experientia, 1968; 24 : 641-752.  Back to cited text no. 8    
9.Quie PG, Mill EL and Holmes B : Molecular events during phagocytosis by human neutrophils, Prog Hematol, 1977; 10 : 193.  Back to cited text no. 9    
10.Stossel TP : Phagocytosis, Amer J Pathol, 1977; 88 : 743:744.  Back to cited text no. 10    
11.Stossel TP : Phagocytosis, Recognition and inges­tion, Semin Hematol, 1975; 12 : 83-116.  Back to cited text no. 11    
12.BabiorBM :Oxygen dependent microbial killing by phagocytes, N Eng J Med, 1978; 298 721-727.  Back to cited text no. 12    
13.Johnston RB Jr : Oxygen metabolism and the microbicidal activity of macrophages, Federal Proc, 1978; 37 : 2759.  Back to cited text no. 13    
14.Ambruso DR, Johnston RB Jr :Defects of phagocyte function, Primary and Secondary Immunodeficiency Disorders, First ed, Editor, Chandra RK :Churchill Livingstone, New York, 1983; p 138-148.  Back to cited text no. 14    
15.Baehner RL, Murramann SK, Davis J et al The role of superoxide anion and hydrogen per­oxide in phagocytosis associated oxidative meta­bolic reactions, J Clin Invest, 1975; 56 : 511­576.  Back to cited text no. 15    
16.Baehner RL and Nathan DG : Quantitative nitroblue tetrazolium test in chronic granulo­matous disease, N Eng J Med, 1961; 278 : 971­-976.  Back to cited text no. 16    
17.Klebanoff SJ : A peroxidase mediated antimi­crobiol system in leukocytes, J Clin Invest. 1967; 46 : 1078 (Abstract).  Back to cited text no. 17    
18.Panton PN and Valentine FCO : Brit J Exp Pathol, 1929; 10 : 257, Quoted by Wilson GS and Miles A : Staphylococcal diseases, in: Princi­ples of Bacteriology, Virology and Immunity, Sixth ed, Editors, Topley WWC and Wilson GS Edward Arnold Publishers, London, 1975; p 1948-1977.  Back to cited text no. 18    
19.Boe J : Acta Dermato-Venereol (Stockh), 1946: 26 : 111, Quoted by Wilson GS and Miles A Staphylococcal diseases, in: Principles of Bacterio­logy, Virology and Immunity, Sixth ed, Editors, Topley WWC and Wilson GS : Edward Arnold Publishers, London, 1975; p 1948-1977.  Back to cited text no. 19    


[Table - 1]


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