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. 1990 May;32(3):195–200. doi: 10.1007/BF01771457

A multicellular tumor spheroid model of cellular immunity against head and neck cancer

Peter G Sacks 1,2, Dorothy L Taylor 1, Tamas Racz 1,3, Tracey Vasey 2, Victon Oke 2, Stimson P Schantz 1,✉
PMCID: PMC11038083  PMID: 2289213

Abstract

A multicellular tumor spheroid (MTS) model for head and neck cancers has been used to examine the immune function of fresh and 6-day interleukin-2(IL-2)-activated peripheral blood lymphocytes (PBL). MTS are individually cultured in the presence of effector cells, and the spheroids' growth is monitored by sizing them under an inverted microscope. Dose/response studies for IL-2 (0–100 U/ml) alone and for fresh unstimulated PBL (0–105 cells/MTS) showed no effects on MTS growth. IL-2-activated PBL (0–105 cells/MTS), in contrast, modulated MTS growth in a multiphasic pattern: MTS growth was unperturbed for the first 3 days and then growth inhibition occurred, followed by MTS disintegration. Histological analysis showed that intact MTS histoarchitecture correlated with unperturbed growth, and increasing cell sloughing and MTS dissolution and replacement by activated PBL correlated with growth inhibition and disintegration. Flow-cytometric sorting of lymphocyte subset populations indicated that it was the Leu19+CD3− cells that produced these growth-modulatory effects. In contrast to the initial LAK cell resistance of MTS, single-cell suspensions demonstrated significant lysis in standard 4-h chromium-release assays. Differences between single cells and MTS suggest a potential for tissue-like organization as a factor in lymphokine-activated killing.

Keywords: Neck Cancer, Peripheral Blood Lymphocyte, Lymphocyte Subset, Multicellular Tumor Spheroid, Subset Population

Footnotes

Supported in part by the First Independent Investigator Award (R29 CA46 251-01) (S. P. S.) of the National Cancer Institute. The Cancer Information Systems Core Facility used in this study was funded under the National Cancer Institute grant CA 16672

References

  • 1.Carlsson J, Nederman T. Tumour spheroid technology in cancer therapy research. Eur J Cancer Clin Oncol. 1989;24:1127. doi: 10.1016/0277-5379(89)90404-5. [DOI] [PubMed] [Google Scholar]
  • 2.Durand RE, Sutherland RM. Effects of intercellular contact on repair of radiation damage. Exp Cell Res. 1972;71:75. doi: 10.1016/0014-4827(72)90265-0. [DOI] [PubMed] [Google Scholar]
  • 3.Grimm EA, Mazumder A, Zhang HZ, Rosenberg SA. Lymphokine-activated killer cell phenomenon: lysis of natural killer-resistant fresh solid tumor cells by interleukin-2-activated autologous human peripheral blood lymphocytes. J Exp Med. 1982;155:1823. doi: 10.1084/jem.155.6.1823. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Herberman RB. Lymphokine-activated killer cell activity. Characteristics of effector cells and their progenitors in blood and spleen. Immunol Today. 1987;8:178. doi: 10.1016/0167-5699(87)90035-1. [DOI] [PubMed] [Google Scholar]
  • 5.Jääskeläinen J, Kalliomäki P, Paetau A, Timonen T Effect of LAK cells against three-dimensional tumor tissue. In vitro study using multi-cellular human glioma spheroids as targets. J Immunol 142: 1036 [PubMed]
  • 6.Lord EM, Burkhardt G. Assessment of in situ host immunity of syngeneic tumors utilizing the multicellular model. Cell Immunol. 1984;85:340. doi: 10.1016/0008-8749(84)90248-x. [DOI] [PubMed] [Google Scholar]
  • 7.Lord EM, Nardella GB, Sutherland RM. The multicellular tumor spheroid model: I. Characterization of the secondary allograft response in sensitized mice. Cell Immunol. 1980;52:119. doi: 10.1016/0008-8749(80)90405-0. [DOI] [PubMed] [Google Scholar]
  • 8.Mickel RA, Kessler DJ, Taylor JMG, Lichtenstein A. Natural killer cell cytotoxicity in the peripheral blood, cervical lymph nodes, and tumor of head and neck cancer patients. Cancer Res. 1988;48:5017. [PubMed] [Google Scholar]
  • 9.Mueller-Klieser W. Multicellular spheroids. A review on cellular aggregates in cancer research. J Cancer Res Clin Oncol. 1987;113:101. doi: 10.1007/BF00391431. [DOI] [PubMed] [Google Scholar]
  • 10.Rupniak HT, Rowlatt C, Lane EB, Steele JG, Trejdosiewicz LK, Laskiewicz B, Povery S, Hill BT. Characteristics of four new human cell lines derived from squamous cell carcinomas of the head and neck. JNCI. 1985;75:621. [PubMed] [Google Scholar]
  • 11.Sacks PG. Growth of head and neck squamous cell carcinoma cell lines as multicell tumor spheroids. In: Wolf GT, Carey TE, editors. Oncology research. Amsterdam: Kugler & Ghedini; 1988. p. 3. [Google Scholar]
  • 12.Sacks PG, Miller MW, Sutherland RM. Response of multicell spheroids to 1 MHz ultrasonic irradiation: cavitation related damage. Radiat Res. 1983;93:545. [PubMed] [Google Scholar]
  • 13.Sacks PG, Parnes SM, Gallick GE, Mansouri Z, Lichtner R, Satya-Prakash KL, Pathak S, Parsons DF. Establishment and characterization of two new squamous cell carcinoma cell lines derived from tumors of the head and neck (1988) Cancer Res. 1988;48:2858. [PubMed] [Google Scholar]
  • 14.Sacks PG, Oke V, Amos B, Vasey T, Lotan R. Modulation of growth, differentiation, and glycoprotein synthesis by β-all-trans retinoic acid in a multicellular tumor spheroid model for squamous carcinoma of the head and neck. Int J Cancer. 1989;44:926. doi: 10.1002/ijc.2910440530. [DOI] [PubMed] [Google Scholar]
  • 15.Sacks PG, Oke V, Vasey T, Lotan R. Retinoic acid inhibition of a head and neck multicellular tumor spheroid model. Head Neck. 1989;11:219. doi: 10.1002/hed.2880110305. [DOI] [PubMed] [Google Scholar]
  • 16.Schantz SP, Goepfert H. Multimodality therapy and distant metastases: The impact of natural killer cell activity. Arch Otolaryngol Head Neck Surg. 1987;113:1207. doi: 10.1001/archotol.1987.01860110073011. [DOI] [PubMed] [Google Scholar]
  • 17.Schantz SP, Brown BW, Lira E, Taylor DL, Beddingfield N. Evidence for the role of natural immunity in the control of metastatic spread of head and neck cancer. Cancer Immunol Immunother. 1987;25:141. doi: 10.1007/BF00199955. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18.Schantz SP, Racz T, Ordonez NG, Terry NA, Taylor DL, Bugis S, Sacks PG. Differential sensitivity of head and neck cancers to non-major histocompatibility restricted killer cell activity. J Surg Res. 1990;48:154. doi: 10.1016/0022-4804(90)90208-j. [DOI] [PubMed] [Google Scholar]
  • 19.Sordat B, MacDonald HR, Lees RK. The multicellular spheroid as a model tumor allograft: III.Morphological and kinetic analysis of spheroid infiltration and destruction. (1980) Transplantation. 1980;29:103. doi: 10.1097/00007890-198002000-00004. [DOI] [PubMed] [Google Scholar]
  • 20.Sutherland RM. Cell and environment interactions in tumor microregions: the multicell spheroid model. Science. 1988;240:177. doi: 10.1126/science.2451290. [DOI] [PubMed] [Google Scholar]
  • 21.Sutherland RM, MacDonald HR, Howell RL. Multicellular spheroids: a new model target for in vitro studies of immunity to solid tumor allografts: brief communication. JNCI. 1977;58:1849. doi: 10.1093/jnci/58.6.1849. [DOI] [PubMed] [Google Scholar]
  • 22.Sutherland RM, Durand RE. Growth and cellular characteristics of multicell spheroids. Recent Results Cancer Res. 1984;95:24. doi: 10.1007/978-3-642-82340-4_2. [DOI] [PubMed] [Google Scholar]
  • 23.Trinchieri G, Perussia B. Human natural killer cells: biologic and pathologic aspects. Lab Invest. 1984;5:489. [PubMed] [Google Scholar]

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