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Objective To investigate the effect of local hyperthermia on the morphology and quantity of Langerhans cells (LCs)at challenged skin sites of a mouse model of contact hypersensitivity.Methods Sixty mice were equally divided into 3 groups to be treated with local hyperthermia (37℃,39℃,41℃and 43℃)for 20 minutes at sensitization sites on the back of mice 3 days before (pre-heat group),concurrently with (concurrent-heat group)or 2 days after(post-heat group)sensitization respectively.Five mice which remained unsensitized and untreated served as the controls.Five days after the sensitization,the mice were challenged on the dorsal surface of right ears.Two days after the elicitation,the right ears were resected and immunohistochemistry was performed to observe the morphology and determine the quantity of LCs at challenged sites.Results With the rise in temperature,the number of LCs in the epidermis of ear skin decreased in pre-heat group(321.83±41.81,251.12±16.29,191.41±28.7,128.33±77.61 per square millimeter at 37 ℃,39 ℃,41℃and 43℃,respectively,P0.05)and in the post-heat group(320.83±113.6,398.33±31.91,437.83±29.78,477.25±86.79 per square millimeter at 37℃,39 ℃,41℃and 43℃,respectively,P<0.01).The dendrites of LCs increased in number and length when the temperature lose from 37 ℃ to 41℃,but slightly declined at 43℃.Conclusions Local hyperthermia at sensitization sites could affect the morphology and density of LCs at challenged sites,and the effect is likely associated with the severity of contact hypersensitivity.
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Objective To investigate the effect of local hyperthermia on the mRNA expression of phosphatidylinositol 3-kinase(P13-K)in Langerhans cells(LCs)transmigrating from HPV-infected skin.Methods Tissue samples were collected from 5 female patients with condyloma acuminatum(CA)and 5 normal human controls.then equally divided into three parts to receive local hyoerthermia treatment at 37℃,42 ℃ and 45℃,respectively,for 30 minutes.After another 12-hour incubation in RPMI 1640 culture medium,transmigrating cells were collected and LCs were purified.Real-time quantitative reverse transcription-PCR was performed to detect the mRNA expressions of P13-K p85a and p110α in purified CD1a~+ LCs from these tissue samples.Results In CD1a~+ LCs transmigrating from normal skin and lesions of CA.the expressions of P13-K p85a and P110α mRNA decreased with the increase of temperature.Atier local hyperthermia treatment of normal skin at 37℃,42℃,45℃,the relative mRNA expression level was 1.00±0.00,0.78±0.13,0.54±0.17,respectively.for P13-K p85α in transmigrating LCs,1.00±0.00,0.80±0.11,0.61±0.12,respectively.for P13-K P110α;there was a significant difierence among the three temperatures in bOth parameters(both P＜0.01).In the case of lesions of CA.the relative mRNA expression level of P13-K p85αand p110α was 1.00±0.00 and 1.00±0.00 under treatment at 37℃,0.20±0.11 and 0.49±0.21 at 42℃.0.1±0.08 and 0.09±0.03 at 45℃.respectively;significant difierence was also noted among the three treatment temperatures(both P＜0.01).The decrease in P13-K P110α mRNA expressions under hyperthermia treatment at 42 ℃ and 45℃ compared with those at 37℃was greater in LCs from CA lesions than that from normal skin.Conclusions Local hyoerthermia could remarkably inhibit the expression of P13-K genes.which may enhance immunity and favor elimination of HPV.
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Objective To observe the morphology, quantity and migration of Langerhans cells in normal or HPV-infected skin treated with localized hyperthermia. Methods Tissue specimens obtained from 16 patients with condyloma accuminatum (CA) and 15 normal controls were divided into three equal parts, and irradiated by a self-made hyperthermia equipment at 37℃, 42℃ and 45℃ respectively for 30 minutes. Immunohistochemistry and flow cytometry were applied to detect the morphology, quantity and migration of Langerhans cells (LCs), respectively, in these treated specimens. Results With a rise in temperature, the number of LCs in epidermis decreased, the dendrites shortened, decreased in number or even disappeared. After exposure to hyperthermia at 37℃, 42℃ and 45℃, the number of LCs was 782.40±114.8, 649.44±119.40 and 510.88±118.64 per square millimeter respectively, in normal tissue, 646.04±135.67, 489.38±118.19 and 387.93±110.15 per square millimeter respectively in HPV-infected skin tissue.The percentage of migratory LCs expressing CD1a was 0.19%±0.18%, 0.89%±0.19% and 1.59%±0.28% in normal skin tissue treated with hyperthermia at 37℃, 42℃ and 45℃ respectively, 0.62%±0.31%,2.31%±0.54% and 6.33%±0.98%, respectively, in HPV-infected skin tissue; the differences were significant among these different temperatures. Furthermore, the migration of LCs from tissue into culture was enhanced by the treatment with hyperthermia. Conclusions Hyperthermia can promote the migration of LCs, and accordingly enhance the antigen presenting effect of these cells in immune response.