Natural killer T cells infiltrate neuroblastomas expressing the chemokine CCL2.
CD1d-restricted Valpha24-Jalpha18-invariant natural killer T cells (iNKTs) are potentially important in tumor immunity. However, little is known about their localization to tumors. We analyzed 98 untreated primary neuroblastomas from patients with metastatic disease (stage 4) for tumor-infiltrating iNKTs using TaqMan((R)) reverse transcription polymerase chain reaction and immunofluorescent microscopy. 52 tumors (53%) contained iNKTs, and oligonucleotide microarray analysis of the iNKT(+) and iNKT(-) tumors revealed that the former expressed higher levels of CCL2/MCP-1, CXCL12/SDF-1, CCL5/RANTES, and CCL21/SLC. Eight tested neuroblastoma cell lines secreted a range of CCL2 (0-21.6 ng/ml), little CXCL12 (</=0.1 ng/ml), and no detectable CCL5 or CCL21. CCR2, the receptor for CCL2, was more frequently expressed by iNKT compared with natural killer and T cells from blood (P < 0.001). Supernatants of neuroblastoma cell lines that produced CCL2 induced in vitro migration of iNKTs from blood of patients and normal adults; this was abrogated by an anti-CCL2 monoclonal antibody. CCL2 expression by tumors was found to inversely correlate with MYCN proto-oncogene amplification and expression (r = 0.5, P < 0.001), and MYCN-high/CCL2-low expression accurately predicted the absence of iNKTs (P < 0.001). In summary, iNKTs migrate toward neuroblastoma cells in a CCL2-dependent manner, preferentially infiltrating MYCN nonamplified tumors that express CCL2.
Metelitsa
Leonid S
LS
Department of Pediatrics, Division of Hematology-Oncology, MS #57, Childrens Hospital Los Angeles, 4650 Sunset Blvd., Los Angeles, CA 90027, USA.
Wu
Hong-Wei
HW
Wang
Hong
H
Yang
Yujun
Y
Warsi
Zamir
Z
Asgharzadeh
Shahab
S
Groshen
Susan
S
Wilson
S Brian
SB
Seeger
Robert C
RC
D000971:Antineoplastic Combined Chemotherapy Protocols / Q000627:therapeutic use; D001483:Base Sequence; D001932:Brain Neoplasms / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000473:pathology / Q000628:therapy; D045744:Cell Line, Tumor; D018932:Chemokine CCL2 / Q000235:genetics* / Q000276:immunology*; D016259:Genes, myc / Q000235:genetics; D015894:Genome, Human; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology*; D009367:Neoplasm Staging; D009447:Neuroblastoma / Q000097:blood / Q000235:genetics* / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D000090063:Proto-Oncogene Mas; D012334:RNA, Neoplasm / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
C111835:CCL2 protein, human; D018932:Chemokine CCL2; C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D012334:RNA, Neoplasm