Table 1.
Based on literature search and supplied product specifications, the authors compared seven mitogens with PHA-M, the traditional mitogen used for lymphocyte stimulation in DCA.
| Order # (Sigma) | Source | Name | Abbreviation | Mitogene activity |
Notes | ||
|---|---|---|---|---|---|---|---|
| Assay | Concentration (µg ml−1) | Reference | |||||
| L9379 | Phytolacca americana | Pokeweed | PW | BrdU, human PBMN 3H-thymidine |
0.03–0.3 2.5 10 0.2/2.0/10.0 |
Sigma product specific(17, 18) | Performed in canine lymphocytes Performed in humans |
| L4391 | Lipopolysaccharide from E. coli | Lipopolysaccharide | LPS | BrdU, human PBMN 3H-thymidine |
5, maximum 62 330 0.2–20 |
Sigma(18, 19) | No mitogenic effect in rhesus monkeys No mitogenic effect in human lymphocytes |
| C0412 |
Canavalia ensiformis, Jack bean |
Concanavalin A | CNA | BrdU, human PBMN 3H-thymidine 3H-thymidine 3H-thymidine |
≤125 7 0.83 5 0.2/2/10 |
Sigma product specific(17–20) | Examined in rhesus monkeys Examined in canine lymphocytes |
| S0149 | Haemolytic streptococci | Strepolysin-O | SLO |
3H-thymidine (maximum incorporation 4–5 d) |
100 50 |
(17, 21) | Permeabilises animal membranes, T lymphocytes targeted, additive effect in combination with other mitogens(17) |
| L8025 | M. amurensis | Haemagglutinating and mitogenic haemagglutinin (MAH & MAM) | LMA |
3H-thymidine (maximum incorporation 3 d) |
MAM = 2 MAH = 20 |
(20) | Agglutinates human blood group 0 erys |
| L9640 | T. vulgaris | Lectin from T. vulgaris | LTV | ≤5 | Sigma | Agglutination activity | |
| L4144 | P. vulgaris, red kidney bean | Phytohemagglutinin | PHA-L | BrdU, human PBMN 3H-thymidine |
≤5 3 |
Sigma product specific(20) | Leukocyte agglutination at ≤1 µg ml−1 |
| Gibco-BRL, 10576-015 | P. vulgaris, red kidney bean | Phytohemagglutinin | PHA-M | 100 µl ml−1 | (3) | Recommended for biodosimetry purposes | |
Additional information from the literature: 15 % homologous serum significantly enhanced (∼2-fold) the response of rhesus lymphocytes to PHA, CNA and PW(19). One study used even 20 % human AB serum(18). Under optimal conditions in rhesus monkeys, CNA (∼2-fold) > PHA ∼ PW(19). With higher concentration mitogenic effect of phytolectins decreasing(19). LPS in rhesus monkey appeared not stimulatory(19) and in human T&B lymphocytes as well(18). SLO in canine lymphocytes has an additive effect in combination with phytolectins (∼2–3-fold), but phytolectins combined do not(17). SLO is a B-cell mitogen requiring T-cell help and phytolectins stimulate T cells(17). PHA stimulates human T & B cells (ratio ∼ 2:1), and PHA ∼ CNA > PW(18). Max. 3H-thymidine incorporation at 4 d of human lymphocyte culture(18).