|Year : 1983 | Volume
| Issue : 5 | Page : 627-628
Immunological study in cases of phlyctenular Kerato-conjunctivities at `T' cell level
D Srivastava, RN Misra, RP Chaturvedi, Jamal Ahmad
State Institute of Ophthamology, M.D. Eye Hospital, Allahabad, India
State Institute of Ophthamology, M.D. Eye Hospital, Allahabad
Source of Support: None, Conflict of Interest: None
|How to cite this article:|
Srivastava D, Misra R N, Chaturvedi R P, Ahmad J. Immunological study in cases of phlyctenular Kerato-conjunctivities at `T' cell level. Indian J Ophthalmol 1983;31:627-8
The delayed hypersensitivity is mediated by sensitized 'T' cells and that there are much better methods to see delayed hypersensitivity in vitro like leukocyte migration inhibition test and lymphocyte transformation test.
So in the present study we have observed delayed hypersensitivity in cases of phlyctenular keratoconjunctivitis against tubercular protein (`T' cell response) and compared it with the Mantoux test on the same patient.
| Materials and Method|| |
The study was carried out at State Institute of Ophthalmology, Allahabad.
1. 10 ml. fresh venous blood was collected from patients of phlyctenular keratoconjunctivitis, defibrinated by shaking with glass beads, transferred to 30 ml. test tube and was mixed with equal volume of 2% gelatin solution in phosphate buffer saline the test tube was kept in incubator at 37°C for 30 minutes at 30° slant position and then for 15 minutes in upright position.
2. Leukocytes from middle buffylayer is taken out and washed three times with Hank's balanced salt solution by centriguging at 400 g. for 5 minutes each time.
3. The suspension of Leukocytes was made in 0.3 ml. of parker 199 media (pH 7.2) containing 10% horse serum. The suspension was drawn into capillary tube of 1 mm uniform internal diameter and 75 mm length. One end of the tubes were sealed with plasticine and were centrifuged at 220 g for 5 minutes to pack leucocytes. Now the tubes were cut at junction of fluid and cellular lavel. The portion. of tubes containing the cells were fixed in migration clamber containing packer 199 media with and without antigen. The antigen was purified protein derivative (PPD) in amount of .2 cc in each chamber.
4. The migration chambers were kept horizontally at 37°C for 16-20 hrs and the areas of leukocyte migration was measured by projecting the migration chamber on screen with the help of slide projector. The area migrated in test chamber (migration chamber with tubercular antigen) were compared with area migrated in control chamber (migration chamber without tuberantigen). The migration index was calculated from following formula. Migration Index
| Observation|| |
The study was conducted on Twenty one cases of phlyctenular kerate-conjunctivitis, of which thirteen cases were Mantoux positive and eight cases Mantoux negative.
Out total no. of cases were twenty one, out of which thirteen cases were Mantoux positive and Eight cases were Mantoux negative.
Out of thirteen cases the leukocyte migration was unaffected in two cases, in three cases unsignificant inhibition was seen i.e. (more than 80% inhibition) and in eight cases significant inhibition was seen. In Mantoux negative cases, in four cases no inhibition was seen while in two cases inhibition was unsignificant and in two cases significant inhibition was seen.
| Discussion|| |
It was observed that out of twenty one cases, in thirteen cases Mantoux test was positive. Out of these Mantoux positive case in 8 cases (61.5%) significant leukocyte migration inhibition was seen. (The inhibition of leukocyte less than 20% is considered insignificant (Bloom B.R. 1971).
Thus it suggests that Mantoux tests is more sensitive in cases of Phlyctenular Keratoconjunctivitis. Which may be due to appreciably less amount of tuberculis which is necessary to produce a skin reaction than is required to stimulate sensitive lymphocytes so that enough migration inhibitary factor (MIF) is produced to bring about inhibition of macrophages (Youmans 1973).
Out of total 8 Mantoux negative cases in two cases (25%) significant inhibition was seen, this can be explained on the basis that due to the internal compartmentalization of sensitized lymphocytes to such a degree in the body that they are not available in the circulation in sufficient numbers to initiate a positive dermal test. (Scholossman . 1971). So both tests should be carried out simultaneously to produce the delayed hypersensitivity against the tubercular protein specially in cases of Phlyctenular conjunctivitis.
| Summary|| |
The study was conducted on twenty one cases phylctenular Kerato-conjunctivitis Leukocytes were separated from blood of these patients. Leukocyte migration inhibition test was done against tubercular protein (PPD), simultaneously Mantoux test was also done.
Out of twenty one cases, thirteen cases were Mantoux positive, out of this thirteen cases in eight cases Leukocyte migration inhibition test was positive i.e. (61.5%).
In eight Mantoux negative cases the leukocyte migration inhibition test was positive in two cases i.e. 25%).
[Figure - 1]
[Table - 1], [Table - 2]