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. 1990 May;31(3):139–145. doi: 10.1007/BF01744727

Fate of intravenously administered rat lymphokine-activated killer cells labeled with different markers

Azzam A Maghazachi 1,✉, Linda Fitzgibbon 1
PMCID: PMC11038838  PMID: 2337903

Abstract

Rat lymphokine-activated killer (LAK) cells, generated by adhering rat splenocytes isolated from the 52% Percoll density fraction to plastic flasks, demonstrate restricted in vivo tissue distribution, localizing in the lungs and liver after 2 h, but redistributing into the liver and spleen 24 h after i.v. administration. However, a different pattern of distribution was observed when this population of LAK cells was labeled with one of four commonly used radioisotopes. For example, LAK cells showed a high distribution into the lungs 30 min after administration when labeled with51Cr,125I-dUrd or111In-oxine, whereas111InCl-labeled LAK cells showed an equal distribution into the blood, lungs and liver at this time. Two hours after administration, cells labeled with111In-oxine showed an equivalent distribution into the lungs and liver, those labeled with125I-dUrd or51Cr showed a high accumulation in the lungs, whereas those labeled with111In-Cl entered more into the liver and blood. The pattern of distribution of111In-Cl- or111In-oxine-labeled cells was confirmed using gamma camera imaging analysis. By 24 h, LAK cells labeled with111InCl,111In-oxine or51Cr distributed in the liver and spleen in variable concentrations. In contrast, cells labeled with125I-dUrd were not detected in any organ tested.

This study was paralleled by monitoring the distribution of LAK cells labeled with Hoechst 33342 (H33342) and analyzed for the presence of fluoresceinated cells in different organs either by flow cytometry analysis, or in frozen section. The data indicate that the distribution pattern of LAK cells labeled with111In-oxine is the closest to the distribution of H33342-labeled cells. Of all the radioisotopes used,125I-dUrd has the most disadvantages and is not recommended for monitoring the in vivo distribution of leukocytes.

Keywords: Freeze Section, Tissue Distribution, Flow Cytometry Analysis, High Accumulation, Gamma Camera

References

  • 1.Ames IH, Gagne GM, Garcia AM, John PA, Scatorchia GM, Tumar RH, McAfee JG. preferential homing of tumor-infiltrating lymphocytes in tumor bearing mice. Cancer Immunol Immunother. 1989;29:93. doi: 10.1007/BF00199283. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2.Brenan M, Parish CR. Intracellular fluorescent labelling of cells for analysis of lymphocyte migration. J Immunol Methods. 1984;74:31. doi: 10.1016/0022-1759(84)90364-8. [DOI] [PubMed] [Google Scholar]
  • 3.Butcher EC, Weissman IL. Direct fluorescent labeling of cells with fluorescein or rhodamine isothiocyanate. I. Technical aspects. J Immunol Methods. 1980;37:97. doi: 10.1016/0022-1759(80)90195-7. [DOI] [PubMed] [Google Scholar]
  • 4.Cheong L, Rich MA, Eidinoff ML. Introduction of the 5-halogenated uracil moiety into deoxyribonucleic acid of mammalian cells in culture. J Biol Chem. 1960;235:1441. [PubMed] [Google Scholar]
  • 5.Chin GW, Cahill RNP. The appearance of fluorescein-labelled lymphocytes in lymph following in vitro or in vivo labeling: the route of lymphocyte recirculation through mesenteric lymph nodes. Immunology. 1984;52:341. [PMC free article] [PubMed] [Google Scholar]
  • 6.Hoffer KG, Hughes WL. Radiotoxicity of intranuclear tritium,125iodine and131iodine. Radiat Res. 1971;47:94. [PubMed] [Google Scholar]
  • 7.Hosokawa M, Sawamura Y, Morikage T, Okada F, Yu Z.-Y., Morikawa K, Itoh K, Kobayashi H. Improved therapeutic effects of interleukin-2 after the accumulation of lymphokine-activated killer cells in tumor tissue of mice previously treated with cyclophosphamide. Cancer Immunol Immunother. 1988;26:250. doi: 10.1007/BF00199937. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Lampidis TJ, Bernal SD, Summerhayes IC, Chen LB. Selective toxicity of rhodamcine 123 in carcinoma cells in vitro. Cancer Res. 1983;43:716. [PubMed] [Google Scholar]
  • 9.Lotze MT, Line BR, Mathisen DJ, Rosenberg SA. The in vivo distribution of autologous human and murine lymphoid cells grown in T cell growth factor (TCGF): implication for the adoptive immunotherapy of tumors. J Immunol. 1980;125:1487. [PubMed] [Google Scholar]
  • 10.Maghazachi AA. In vivo chemotaxis and chemokinesis activities of IFNγ, IFNα and IL-2 for rat lymphokine-activated killer cells. Nat Immun Cell Growth Regul. 1989;8:130. [Google Scholar]
  • 11.Maghazachi AA, Herberman RB, Vujanovic NL, Hiserodt JC. In vivo distribution and tissue localization of highly purified rat lymphokine-activated killer (LAK) cells. Cell Immunol. 1988;15:179. doi: 10.1016/0008-8749(88)90172-4. [DOI] [PubMed] [Google Scholar]
  • 12.Maghazachi AA, Goldfarb RH, Herberman RB. Influence of T cells on the expression of lymphokine-activated killer cell activity and in vivo tissue distribution. J Immunol. 1988;141:4039. [PubMed] [Google Scholar]
  • 13.Maghazachi AA, Goldfarb RH, Kitson RP, Hiserodt JC, Giffen CA, Herberman RB. Interleukin-2 and killer cells in cancer. Florida: CRC; 1989. In vivo tissue distribution of interleukin-2 activated cells; p. 259. [Google Scholar]
  • 14.Maghazachi AA, Goldfarb RH, Herberman RB. Natural killer cells and host defense. Basel: Karger; 1989. Effect of carbohydrates on the in vivo migration of purified LAK cells; p. 242. [Google Scholar]
  • 15.Marcus CS. The status of indium-111 oxine leukocyte imaging studies. None invasive medical imaging. 1984;1:213. [Google Scholar]
  • 16.Mathias AP, Fisher GA, Prusoff WH. Inhibition of the growth of mouse leukemia cells in culture by 5 iododeoxyuridine. Biochim Biophys Acta. 1959;36:560. doi: 10.1016/0006-3002(59)90209-4. [DOI] [PubMed] [Google Scholar]
  • 17.Mazumdar A, Rosenberg SA. Successful immunotherapy of natural killer-resistant established pulmonary melanoma metastases by the intravenous adoptive transfer of syngeneic lymphocytes activated in vitro by interleukin 2. J Exp Med. 1984;159:495. doi: 10.1084/jem.159.2.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Rannie GH, Donald KJ. Estimation of the migration of thoracic duct lymphocytes to non-lymphoid tissues. A comparison of the distribution of radioactivity at intervals following i. v. transfusion of cells labeled with3H,14C,35S,99mTC,125I and51Cr in the rat. Cell Tissue Kinet. 1977;10:523. [PubMed] [Google Scholar]
  • 19.Rodolfo M, Salvi C, Parmiani G. Influence of the donors' clinical status on in vitro and in vivo tumor cytotoxic activation of interleukin-2-exposed lymphocytes and their circulation in different organs. Cancer Immunol Immunother. 1989;28:136. doi: 10.1007/BF00199114. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Rosenberg SA, Lotze MT. Cancer immunotherapy using interleukin-2 and interleukin-2-activated lymphocytes. Annu Rev Immunol. 1986;4:681. doi: 10.1146/annurev.iy.04.040186.003341. [DOI] [PubMed] [Google Scholar]
  • 21.Rosenberg SA, Mule JJ, Spiess PL, Reichert CM, Schwarz SL. Regression of established pulmonary metastases and subcutaneous tumor mediated by the systemic administration of high-dose recombinant interleukin-2. J Exp Med. 1985;161:1169. doi: 10.1084/jem.161.5.1169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Rosenberg SA, Lotze MT, Muul LM, Chang AE, Avis FP, Leitman S, Lineham WM, Robertson CN, Lee RE, Rubin JT, Seipp CA, Simpson CG, White DE. A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high-dose interleukin-2 alone. N Engl J Med. 1987;316:889. doi: 10.1056/NEJM198704093161501. [DOI] [PubMed] [Google Scholar]
  • 23.Sayle BA, Balachandran S, Rogers CA. Indium-111 chloride imaging in patients with suspected abscesses: concise communication. J Nucl Med. 1983;24:1114. [PubMed] [Google Scholar]
  • 24.Sprent J. Fate of H2-activated T lymphocytes in syngeneic hosts. I Fate in lymphoid tissues and intestines traced with3H-thymidine,125I-deoxyuridine and51chromium. Cell Immunol. 1976;21:278. doi: 10.1016/0008-8749(76)90057-5. [DOI] [PubMed] [Google Scholar]
  • 25.Takai N, Tanaka R, Yoshida S, Hara N, Saito T. In vivo and in vitro effect of adoptive immunotherapy of experimental murine brain tumors using lymphokine-activated killer cells. Cancer Res. 1988;48:2047. [PubMed] [Google Scholar]
  • 26.Vujanovic NL, Herberman RB, Maghazachi AA, Hiserodt JC. Lymphokine-activated killer cells in rats. III. A simple method for the purification of large granular lymphocytes and their rapid expansion and conversion into lymphokine activated killer cells (large granular lymphocytes) J Exp Med. 1988;167:15. doi: 10.1084/jem.167.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]

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