Characterization of functional neurotensin receptors on human lymphocytes.
BACKGROUND
Neurotensin, a tridecapeptide, regulates gut motility, secretion, and mucosal growth; an immunomodulatory role for neurotensin has been postulated but not clearly defined. The purpose of this study was to determine whether neurotensin receptors (NTR) are present on peripheral blood lymphocytes (PBLs) and to characterize binding, functional, and molecular properties.


METHODS
Iodine 125 labeled-neurotensin binding was determined for both human PBLs and the T-cell lines, Molt-4 and Jurkat, by Scatchard analysis. To analyze functional capacity of the NTR, PBLs were cultured in the presence of phytohemagglutinin (1 microgram/ml) with or without neurotensin and harvested at 48 hours after an 8-hour pulse of tritiated thymidine. For molecular analyses, RNA extracted from PBLs and T-cell lines was analyzed by either Northern hybridization with a labeled NTR cDNA or ribonuclease protection with an antisense human NTR probe.


RESULTS
Scatchard analyses of neurotensin binding to human PBLs and MOLT-4 showed two classes of binding sites with different affinities. Incubation of phytohemagglutinin-stimulated PBLs with neurotensin significantly enhanced proliferation. Northern hybridization showed mRNA of the authentic NTR or a receptor subtype of close homology in PBLs and the T-cell lines; ribonuclease protection analysis identified the authentic human NTR in the MOLT-4 cell line.


CONCLUSIONS
Using a combination of molecular techniques and Scatchard analysis, we have demonstrated the presence of a cell surface NTR with a high affinity for neurotensin on human PBLS and the MOLT-4 cell line. The functional role of NTR has been established by enhanced proliferation with addition of neurotensin. These data provide further evidence for a link between neurotensin and the immune system and suggest that neurotensin may play an important regulatory role in gut mucosal immune responses in vivo.
Evers
B M
BM
Department of Surgery, University of Texas Medical Branch, Galveston 77555-0533.
Bold
R J
RJ
Ehrenfried
J A
JA
Li
J
J
Townsend
C M
CM
Klimpel
G R
GR
D015152:Blotting, Northern; D002460:Cell Line; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects; D008214:Lymphocytes / Q000737:chemistry*; D009496:Neurotensin / Q000378:metabolism / Q000494:pharmacology; D018028:Receptors, Neurotensin / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D012260:Ribonucleases / Q000494:pharmacology
D018028:Receptors, Neurotensin; D009496:Neurotensin; D012260:Ribonucleases