|Year : 1993 | Volume
| Issue : 4 | Page : 176-183
Basophil leucocyte response in leprosy with lepromin-A
RP Okhandiar, HS Sharma
R P Okhandiar
Source of Support: None, Conflict of Interest: None
Study of 'Basophil Leucocyte Response' in two polar groups of leprosy patients (tuberculoid and lepromatous) was undertaken employing Lepromin-A (Armadillo) by the method of Open Window Technique of Rebuck. Significant infiltration of Basophils at the test side is demonstrated both at 48 hours and at 72 hours in lepromatous leprosy as compared to the control groups i.e., in placebo and tuberculin positive healthy volunteers as well as against tuberculoid leprosy group (p value being less than 0.05 and 0.001). The basophils also go into degeneration and degranulation at 72 hours. .
The role of basophils in the different types of allergic reactions is discussed in context of above finding. The possible effect of high infiltration of basophils in lepromatous leprosy with its subsequent release of histamine and other factors by degranulation, and its possible role in immuno-regulation through specialised suppressor cells is postulated in the light of available data.
|How to cite this article:|
Okhandiar R P, Sharma H S. Basophil leucocyte response in leprosy with lepromin-A. Indian J Dermatol Venereol Leprol 1993;59:176-83
|How to cite this URL:|
Okhandiar R P, Sharma H S. Basophil leucocyte response in leprosy with lepromin-A. Indian J Dermatol Venereol Leprol [serial online] 1993 [cited 2020 Jun 1];59:176-83. Available from: http://www.ijdvl.com/text.asp?1993/59/4/176/3923
| Introduction|| |
Leprosy is one of those diseases where inspite of extensive studies, the precise nature of the immune defect is still not yet fully understood, especially nature of specific depression of cell- mediated immunity in lepromatous leprosy, as evidenced by negative delayed hypersensitivity reaction.
Although T-lymphocytes and monocyte-macrophage system are the principal essential participants in delayed hypersensitivity skin reaction, it has been noted that products of B-cells, eosinophils, mast cells/basophils may also be involved in varying degrees.
During last one and half decade interest has been created on the role of basophils in delayed hypersensitivity reaction as a result of the work of Dvorak and his colleagues , who demonstrated their participation in such reaction by degranulation and release of histamine. Basophils are found immediately under the epidermis in most delayed hypersensitivity reactions especially in allergic contact dermatitis and Jones - Mote reactions , Fulton and Derbes  also demonstrated the presence of basophils in variable amount in dermal exudate in positive tuberculin skin test reaction. In order to determine, whether similar processes might be involved in the delayed hypersensitivity reaction in leprosy with Lepromin - A (Lepromin prepared from tissue of Armadillo infected with M. Leprae, containing 40 million dead bacilli per ml of the emulsion) the present study has been undertaken using the skin-window technic developed by Rebuck (Fulton and Derbes). 
| Materials and Methods|| |
Thirty seven patients of leprosy (twenty tuberculoid and seventeen lepromatous) age - group between l2 to 40, eleven females and rest males, were selected for this study. Only patients of two polar types i.e., tuberculoid and lepromatous based on the criteria laid down by Indian classification were chosen. Eleven tuberculin positive volunteers between the age group of 16 - 30 years with no evidence of active tuberculosis served as controls.
The following protocol was worked out. On the first day, either 5 T.U. of Old tuberculin, or 0.2 ml of Lepromin-A, or 0.2 ml of Phenol-saline (0.5% phenol in normal saline) was injected intradermally in the middle third of the volar surface of the forearm using 0.1 ml tuberculin syringe and a 26 gauge hypodermic needle.
Twenty-four hours later, on second day a 15 mm diameter (approx.) area of epidermis of the resulting reaction was scraped off with sterile scalpel blade. An area of about 15 mm diameter was scraped down to the rate ridges. Care was taken that the lesions did not bleed. Following the removal of the skin, sterile round glass cover-slips (18 mm size) were placed over the lesions. Sterile gauge pads were put over the glass to keep the coverslips from breaking. Finally the material was taped in place with leucoplast and the whole area was bandaged.
Four hours later i.e., at 28 hours after the initial injection, the first coverslip was removed and another was put in its place. A similar exchange was done at 48 hours and at 72 hours.
The cover-slips were stained with Leishman's stain (G.T. Gurr, London) and mounted on a microscope slide with DPX. On each slide two hundred cells were counted in the area that yielded the highest aggregation of cells. The cells were classified in four groups:
(i) Neutrophilic granulocytes.
(ii) Mononuclear cells (lymphocytes, monocytes, macrophages)
(iii) Eosinophilic granulocytes.
(iv) Basophilic granulocytes.
| Results|| |
The findings have been depicted in the micro-photographs and tables. [Figure - 1] shows that at 28 hours in leprosy (both tuberculoid or lepromatous) there is an exudate consisting of 80-90 percent of neutrophilic granulocytes. It is also true for tuberculin positive group as demonstrated by Fulton & Derbes  and same is the case of phenol-saline group [Table - 1].
At 48 hours the character of exudate is changed [Figure - 2]. No longer the neutrophils are the predominant cells. Mononuclear cells have appeared in large number, as well as a few eosinophils. But the most significant change is the Litlux of basophils and the differential count of cell exudate shows [Table - 2] that the influx is more in lepromatous leprosy than that in tuberculoid leprosy or in tuberculin positive group. Even in tuberculoid leprosy there appears to be more basophils than that in tuberculin positive group. In phenol-saline group, however there is no influx of basophil at all .
At 72 hours, the basophils have become hypertrophic and distorted. Most of the basophils have degranulated. Quite a few mononuclear cells have taken up the granules [Figure - 3][Figure - 4]. Again the differential count shows [Table - 3] that the influx of basophils is highest in lepromatous group and gradually decreasing in tuberculoid and tuberculin positive group and almost absent in phenol-saline group, as has been observed at 48 hours.
In [Table - 4] the influx of basophils on lepromatous group has been compared statistically to other groups employing Student's t-test. The influx does not appear to be significant statistically at 28 hours with any of the other groups p value being more than 0.05.
At 48 hours and at 72 hours however the influx of basophils in lepromatous group appears to be highly significant as compared to phenol-saline group or tuberculin positive group or even tuberculoid leprosy group, the p value being less than 0.001 in most of the cases.
| Comments|| |
Basophils leucocytes (and/or mast cells) exhibit chemotaxis and phagocytic activity but their physiologic role is played by discharge of the contents of the specific granules to the exterior.  Probably, the human basophils contain all the blood histamine localised in its granules. Other than histamine, the granules also contain heparin, 5-hydroxytryptamine, eosinophil and neutrophil chemotactic factors, platelet activating factors besides other factors having known and unknown physiologic role.
Basically the basophils (and/or mast cells) are well-established for their role in IgE mediated Type I hypersensitivity reaction. The interaction between the antigen and the IgE attached to the surface of basophils initiate a sequence of reactions furthering imbalance of cyclic neucleotides (i.e., cyclic AMP and cyclic GMP) which leads to degranulation and release of histamine and other substances.
Although the relevance of IgE and basophils/mast cells to Type I type of hypersensitivity reaction is well known, it must not be forgotten that a wide variety of agents may act to degranulate human basophils like radio-contrast media, opiates, curare, dextran, irradiation etc.  Rather than diminish the role of basophils/mast cell in immunologic diseases, this fact, instead, expands the relevance of this cell and its many mediators to other pathophysiologic states independent of antigen or antibody.
Several studies have shown increased histamine level at the sites of contact allergic reactions. , Basophils in the blister fluid and the skin window exudate of contact allergic reaction sites have been well documented. , Dvorak et al , think that their presence can be explained by the co-existence of delayed hypersensitivity reaction of Jone's Mote Type together with conventional "Lymphocyte- macrophage mediated" delayed hypersensitivity.
Now this 'Jone's Mote Reaction" or so-called 11 cutaneous basophil hypersensitivity" is another interesting aspect of immune reaction. These transitory reactions induced by antigen with or without Freund's incomplete adjuvant (as against antigen with Freund's complete adjuvant) have been shown to be the result of more intense immunoregulation by specifically induced suppressor cells by Turk et all  and Ota et al 1 in a series of studies using cyclophosphamide to eliminate suppressor cell precursors.
How this regulatory mechanism works in the periphery is still not clearly understood, but it has been proposed by Askenase  that B- cell produce IgG l antibody locally which acts through the basophils which then produce vaso-active amines locally and dampen the delayed hypersensitivity reaction. Such type of immuno-regulatory process by suppressor cells has been demonstrated in contact sensitivity to simple chemical sensitizer like DNCB and picryl chloride by many workers including Chase  and Turk et al  Askenase et al  have further elucidated the role of these suppressor cells which they name "histamine dependent suppressor cells", in enhancing melanoma tumor growth in a series of experiments using animal model.
What would be the role of basophils in the delayed sensitivity reaction in leprosy with Lepromin - A is more a matter of speculation. Increase in number of mast cells have also been observed in human and murine leprosy.  Kumar et al  demonstrated increased level of serotonin in the blood of lepromatous patient, significance of which remains to be determined. In a comprehensive review on the subject of possible immune-depression in leprosy, Turk & Curtis  feel that the failure could lie at any level starting from monocyte-macrophage system to at T-cell level. They have also laid stress on the role by suppressor cells of specific immuno-regulation, a fact emphasised by Mehra et al . Whether these specialized suppressor cells are "histamine dependent" as envisaged by Nordlund and Askenase  or bear H 2 type histamine receptors (Shearer et al  as quoted by Nath ) or induced by preferential stimulation to large mycobacterial antigen as postulated by Turk et a1  shall remain a matter of future investigation. And thus the present study demonstrating significant basophil infiltration in lepromatous leprosy as compared to tuberculoid leprosy or controls might throw some light to this particular branch of research.
| Acknowledgement|| |
The authors thank Dr S K Noordeen, WHO, Geneva for liberal supply of Lepromin - A.
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[Figure - 1], [Figure - 2], [Figure - 3], [Figure - 4]
[Table - 1], [Table - 2], [Table - 3], [Table - 4]