title: Immunomodulatory effects of low-intensity near-infrared laser irradiation on contact hypersensitivity reaction.
abstract: OBJECTIVE
  Contact hypersensitivity (CHS) reaction is a useful model for studying the skin immune system and inflammatory reactions in the skin. In this study, an experimental model of CHS reaction was employed to assess immunomodulatory effects of near-infrared (near-IR) low-intensity laser (LIL) irradiation, which is used as adjuvant therapy in dermatology, physical medicine, rheumatology, etc., because of its declared anti-inflammatory, biostimulative and analgesic effects.
  
  
  METHODS
  The effects of near-IR LIL irradiation (lambda=904 nm, irradiance 60 mW/cm2, fluence 3.6 J/cm2) on CHS reaction to 1-chloro-2,4-dinitrochlorobenzene (DNCB) in Albino Oxford rats were examined by irradiating experimental groups of animals before the induction phase of CHS reaction, while nonirradiated animals and animals that received vehicle instead of hapten served as controls. Ear-swelling assay, histopathological examination of H&E preparations of ear skin, computer-assisted image analysis of dermal infiltrate, ear skin organ culture with the determination of cutaneous production of tumour necrosis factor-alpha (by ELISA assay) and nitric oxide (by Griess' assay) were used for measuring the effects of LIL in the elicitation phase of CHS reaction. Cellularity, dendritic cell content, flow cytometry and proliferation assays (spontaneous and in the presence of IL-2 and concanavalin A) of the draining lymph node cells (DLNC) were performed for the assessment of LIL irradiation effects in the induction phase.
  
  
  RESULTS
  In the irradiated group of animals, ear swelling was significantly diminished compared to control animals (101+/-11.5% vs. 58+/-11.6%, P<0.01). This was accompanied by a highly significant decrease in the density of dermal infiltrate (22+/-0.81 vs. 14.2+/-1.75 cells per unit area, P<0.01) and a significant decrease in nitrite levels in the medium conditioned by organ-cultured ear skin (17.63+/-1.91 vs. 3.16+/-1.69 microM NaNO2; P<0.01), while TNF-alpha concentration was not changed. Cellularity and dendritic cell content in DLNC population, as well as the expression of TCR-alpha, CD4, CD8 and CD25, were not changed between irradiated and nonirradiated animals. Proliferation rates of DLNC cultured for 72 h were significantly lower in irradiated animals (17.3+/-4.1 vs. 13.9+/-0.9 x 103 c.p.m.; P<0.01). In cultures of DLNC with added rIL-2 or 0.5 microg/ml of concanavalin A, proliferation rates were also significantly decreased in irradiated animals (34.7+/-3.5 vs. 31.2+/-2. c.p.m. in IL-2-supplemented culture, P<0.01; 70.9+/-6.4 vs. 58.3+/-9.1 x 103 c.p.m. in concanavalin A-supplemented culture, P<0.01). However, this effect was overcome in the presence of the higher concentration of concanavalin A (2.5 microg/ml) (nonirradiated 38.7+/-3.1, irradiated 123.1+/-7.3 x 103 c.p.m., P<0.01).
  
  
  CONCLUSIONS
  LIL irradiation showed a systemic immunomodulatory effect on CHS reaction to DNCB in rats. Decreased ear swelling observed in the elicitation phase was associated with diminished proliferative responses of the DLNC in the induction phase of CHS reaction. Further experimental work is needed to examine the possible mechanisms of these effects.
authors.affiliation: Department of Dermatology and Institute for Medical Research, Military Medical Academy, Belgrade, Serbia and Montenegro.
authors.forename: Lidija
authors.initials: L
authors.lastname: Kandolf-Sekulovic
authors.forename: Milena
authors.initials: M
authors.lastname: Kataranovski
authors.forename: Milos D
authors.initials: MD
authors.lastname: Pavlovic
mesh_terms: D000818:Animals; D017449:Dermatitis, Allergic Contact / Q000276:immunology / Q000532:radiotherapy*; D004137:Dinitrochlorobenzene; D004431:Ear, External; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D007259:Infrared Rays; D008198:Lymph Nodes / Q000166:cytology; D008297:Male; D010789:Phototherapy / Q000379:methods*; D051381:Rats; D011919:Rats, Inbred Strains; D018709:Statistics, Nonparametric
chemical_list: D004137:Dinitrochlorobenzene