Mishell–Dutton culture

Mishell–Dutton culture (also called the Mishell–Dutton technique) is an in-vitro primary antibody response system using dissociated mouse spleen cells. First reported in 1966–1967 by Robert I. Mishell and Richard W. Dutton, it enabled reliable in-vitro immunization and enumeration of antibody-forming cells via plaque-forming cell (PFC) assays, facilitating studies of B–T cell cooperation and accessory cell requirements.[1][2]

Background

The culture's quantitative readout relied on single-cell antibody detection methods: the PFC assay of Jerne and Nordin and later refinements by Cunningham and Szenberg.[3][4]

Methods manual

A widely used bench manual that compiled many related procedures is Selected Methods in Cellular Immunology (1980), edited by Barbara B. Mishell and Stanley M. Shiigi.[5]

Impact

The system was central to dissecting cellular cooperation in primary antibody responses, including evidence for two-cell requirements and antigen-presenting cell–dependent clustering in vitro.[6][7] Subsequent optimizations—notably the addition of 2-mercaptoethanol to culture media—further enhanced response magnitude and reproducibility.[8]

See also

References

  1. ^ Mishell, Robert I.; Dutton, Richard W. (1966). "Immunization of normal mouse spleen cell suspensions in vitro". Science. 153 (3739): 1004–1006. doi:10.1126/science.153.3739.1004. PMID 5917547.
  2. ^ Mishell, Robert I.; Dutton, Richard W. (1967). "Immunization of dissociated spleen cell cultures from normal mice". The Journal of Experimental Medicine. 126 (3): 423–442. doi:10.1084/jem.126.3.423. PMC 2138377. PMID 6034749.
  3. ^ Jerne, Niels K.; Nordin, Albert A. (1963). "Plaque formation in agar by single antibody-producing cells". Science. 140 (3565): 405. doi:10.1126/science.140.3565.405. PMID 17815805.
  4. ^ Cunningham, A. J.; Szenberg, A. (1968). "Further improvements in the plaque technique for detecting single antibody-forming cells". Immunology. 14 (4): 599–600. PMC 1409394. PMID 5653817.
  5. ^ Mishell, Barbara B.; Shiigi, Stanley M. (1980). Selected Methods in Cellular Immunology. San Francisco: W. H. Freeman. ISBN 0-7167-1106-0.
  6. ^ Mosier, D. E. (1967). "A requirement for two cell types for antibody formation in vitro". Science. 158 (3808): 1573–1575. doi:10.1126/science.158.3808.1573. PMID 6060362.
  7. ^ Mosier, D. E. (1969). "Cell interactions in the primary immune response in vitro: A requirement for specific cell clusters". The Journal of Experimental Medicine. 129 (2): 351–362. doi:10.1084/jem.129.2.351. PMC 2138600. PMID 5767312.
  8. ^ Click, R. E.; Benck, L.; Alter, B. J. (1972). "Enhancement of antibody synthesis in vitro by mercaptoethanol". Cellular Immunology. 3 (1): 156–160. doi:10.1016/0008-8749(72)90237-7. PMID 5061825.
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