Characterization of the T-dependent antibody response (TDAR) to keyhole limpet hemocyanin (KLH) in the Göttingen minipig

Figures & data

Table 1. Treatment groups.

			KLH challenge (mg/animal)		Number of animals
Group number	Treatment	Dose level (mg/kg/day)	1°	2°	M	F
1	Saline	0	2	2	4	1
2	Saline	0	10	2	4	1
3	CPS	2.5	10	2	2	0
4	CPS	12.5	10	2	2	0
5	CPS	25	10	2	2	0
6	CPS	50	2	2	2	1
7	CPS	50	10	2	2	1

Animals were treated with saline or the assigned dose of CPS once daily on Days 0–4 and on Days 14–18. One animal in Group 5 did not receive treatment with CPS on Days 14–18. KLH immunizations were given on Day 0 and on Day 14 (following blood collection).

M, male; F, female.

Figure 1. Primary IgM, primary IgG, and secondary IgG anti-KLH antibody responses of Göttingen Minipigs® immunized with 2 mg KLH. Saline-treated minipigs (males: animals A–D; female: animal E) were immunized with 2 mg KLH by IV injection via ear vein on Day 0. On Day 14, all animals received a secondary immunization of 2 mg KLH by IV injection via ear vein. Primary antibody levels (left panel for each animal) were evaluated in serum samples obtained on Days -6, 5, 6, 7, 8, 9, 12, 13, and 14. Secondary IgG antibody levels (right panel for each animal) were evaluated in serum samples obtained on Days 19, 20, 21, 22, and 23. Results are presented as mean ± SE in terms of log₂ (titer). Dotted horizontal line: baseline (i.e. Day -6) primary IgM antibody levels. Solid horizontal line: baseline (i.e. Day -6) primary IgG antibody levels.

Figure 2. Primary IgM, primary IgG, and secondary IgG anti-KLH antibody responses of Göttingen Minipigs® immunized with 10 mg KLH. Saline-treated minipigs (males: animals A–D; female: animal E) were immunized with 10 mg KLH by IV injection via ear vein on Day 0. On Day 14, all animals received a secondary immunization of 2 mg KLH by IV injection via ear vein. Primary antibody levels (left panel for each animal) were evaluated in serum samples obtained on Days -6, 5, 6, 7, 8, 9, 12, 13, and 14. Secondary IgG antibody levels (right panel for each animal) were evaluated in serum samples obtained on Days 19, 20, 21, 22, and 23. Results are presented as mean ± SE in terms of log₂ (titer). Dotted horizontal line: baseline (i.e. Day -6) primary IgM antibody levels. Solid horizontal line: baseline (i.e. Day -6) primary IgG antibody levels.

Figure 3. Primary IgM, primary IgG, and secondary IgG anti-KLH antibody responses of Göttingen Minipigs® treated with cyclophosphamide. Male minipigs were treated with 2.5 (animals A and B), 12.5 (animals C and D), or 25 mg CPS/kg/day (animals E and F) daily on Days 0–4 and on Days 14–18. Animal E received no CPS treatment on Days 14–18. All animals were immunized with 10 mg KLH by IV injection via ear vein on Day 0. On Day 14, all animals received a secondary immunization of 2 mg KLH by IV injection via ear vein. Primary antibody levels (left panel for each animal) were evaluated in serum samples obtained on Days -6, 5, 6, 7, 8, 9, 12, 13, and 14. Secondary IgG antibody levels (right panel for each animal) were evaluated in serum samples obtained on Days 19, 20, 21, 22, and 23. Results are presented as mean ± SE in terms of log₂ (titer). Dotted horizontal line: baseline (i.e., Day -6) primary IgM antibody levels. Solid horizontal line: baseline (i.e., Day -6) primary IgG antibody levels.

Table 2. Optimum immunization and serum sampling parameters for assessing the TDAR in Göttingen Minipigs®.

Parameter	Optimal result
Primary Immunization	10 mg KLH on Day 0
Primary IgM Antibody Levels	Collect serum daily on Days 6, 7, 8, and 9
Primary IgG Antibody Levels	Collect serum daily on Days 12, 13, and 14
Secondary Immunization	2 mg KLH on Day 14
Secondary IgG Antibody Levels	Collect serum daily on Days 20, 21, and 22