title: Immunoregulatory effects of freeze injured whole tumour cells on human dendritic cells using an in vitro cryotherapy model.
abstract: Tumour cryotherapy has been described as both immunostimulatory and immunoinhibitory in previous studies. However, previous studies have not accurately reproduced the precise conditions of current clinical cryotherapy. The objective of this study is to assess the immunological effects of cryotreated whole tumour cells on dendritic cells (DC) maturation and function using an in vitro model. Prostate cancer cells were cooled using Endocare cryo-system to mimic temperatures achieved during clinical cryotherapy. Human DC were prepared from cluster of differentiation (CD) 14 monocytes and matured with lipopolysaccharide (LPS). Cryotreated cancer cells were added to DC on day 3. On day 7, DC were harvested and phenotyped. Cytokine gene expression was assessed using real time quantitative polymerase chain reaction (RT-PCR). Functional activity of DC was assessed in allogenic mixed lymphocyte reaction (MLR) and the molecular changes using gene microarray technology. There was statistically significant upregulation of costimulatory molecules and maturation markers (CD86, CD83, CD80 and CL II) in DC loaded with cryotreated whole tumour cells compared to both control DC and DC matured with LPS (P < 0.001). There was a significant increase in stimulatory cytokines gene expression (IL-2, IL-12, IL-15, IL-18 and IFN-γ). However, IL-10 and TGF-β expression reduced significantly. The effect of different freezing temperature was equal. cDNA microarray analysis showed upregulation of interleukin 1 (IL-1) and cycline dependent kinase inhibitor 1A (CDKN1A (p21) and downregulation of Caspase 8 and BCL2. Overall, our findings suggest that the effect of cryotherapy is generally stimulatory to DC which may enhance anti-tumour effects. Therefore, the combination of cryotherapy and DC vaccine may represent a novel method to increase the efficacy of cryotherapy especially at the peripheral zones of the prostate where cells are exposed to sub-lethal temperature.
authors.affiliation: Postgraduate Medical School, University of Surrey, Guildford, Surrey GU27WG, UK. ms18273@gmail.com
authors.forename: Mohamed
authors.initials: M
authors.lastname: Ismail
authors.forename: Richard
authors.initials: R
authors.lastname: Morgan
authors.forename: Kevin
authors.initials: K
authors.lastname: Harrington
authors.forename: John
authors.initials: J
authors.lastname: Davies
authors.forename: Hardev
authors.initials: H
authors.lastname: Pandha
mesh_terms: D015703:Antigens, CD / Q000096:biosynthesis; D018122:B7-1 Antigen / Q000096:biosynthesis; D051940:B7-2 Antigen / Q000096:biosynthesis; D045744:Cell Line, Tumor; D017679:Cryotherapy / Q000379:methods*; D003713:Dendritic Cells / Q000276:immunology*; D005615:Freezing; D020869:Gene Expression Profiling; D006801:Humans; D007136:Immunoglobulins / Q000096:biosynthesis; D007378:Interleukins / Q000096:biosynthesis; D008297:Male; D008562:Membrane Glycoproteins / Q000096:biosynthesis; D011471:Prostatic Neoplasms / Q000628:therapy*; D015854:Up-Regulation; D000097802:CD83 Antigen
chemical_list: D015703:Antigens, CD; D018122:B7-1 Antigen; D051940:B7-2 Antigen; D007136:Immunoglobulins; D007378:Interleukins; D008562:Membrane Glycoproteins; D000097802:CD83 Antigen; C497964:CD86 protein, human