Phenotype of lymphocytes in the small and large intestine by flow cytometrya
| All Values Mean % (SD) . | IELs . | . | . | . | LPLs . | . | . | . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Age . | Small intestine . | . | Large intestine . | . | Small intestine . | . | Large intestine . | . | ||||||
| . | <9 mo . | >9 mo . | <9 mo . | >9 mo . | <9 mo . | >9 mo . | <9 mo . | >9 mo . | ||||||
| Samples (n) | 6 | 3 | 13 | 8 | 6 | 3 | 13 | 8 | ||||||
| % Lymphocytes CD3+ Mean (SD) | 11.4 (10.5) | 28.0 (3.1) | 21.0 (15.7) | 33.8 (13.6) | 28.03 (18.0) | 64.27 (0.5) | 51.59 (27.0) | 54.18 (18.1) | ||||||
| Samples (n) | 4 | 1 | 7 | 4 | 4 | 1 | 7 | 4 | ||||||
| 3+4+ | 18.2 (24.1) | 46.9 (NA) | 20.2 (16.5) | 41.4 (24.5) | 52.5 (14.9) | 57.7 (NA) | 57.6 (11.9) | 51.5 (9.9) | ||||||
| 3+8+ | 20.2 (15.5) | 26.9 (NA) | 34 (23.1) | 31.3 (10.6) | 19.9 (8.4) | 24.3 (NA) | 24.6 (6.7) | 27.7 (9.3) | ||||||
| 3+4+8+ | 3.3 (2.3) | 3 (NA) | 1.9 (1.8) | 3.5 (4.2) | 4.7 (3.9) | 1.7 (NA) | 2.2 (2.1) | 6.4 (5.2) | ||||||
| 3+4−8− | 58.3 (36.8) | 23.2 (NA) | 43.9 (33.9) | 23.9 (15.1) | 22.9 (14.0) | 16.3 (NA) | 15.6 (10.0) | 14.5 (3.8) | ||||||
| Samples (n) | 1 | 1 | 3 | 2 | 1 | 1 | 3 | 2 | ||||||
| 3−7+2− | 8.7 (NA) | 0.5 (NA) | 9.3 (7.6) | 3.9 (5.2) | 5.9 (NA) | 0.8 (NA) | 5.7 (3.6) | 1.9 (2.3) | ||||||
| 3−7+2+ | 5.3 (NA) | 0.5 (NA) | 6.2 (3.2) | 3.4 (3.4) | 6.2 (NA) | 1.2 (NA) | 8.2 (4.4) | 6.0 (6.9) | ||||||
| 3+7+2− | 5.6 (NA) | 6.5 (NA) | 2.5 (0.5) | 2.0 (0.7) | 8.2 (NA) | 8.9 (NA) | 6.4 (0.8) | 3.5 (1.8) | ||||||
| 3+7−2+ | 13.5 (NA) | 15.7 (NA) | 23.7 (14.2) | 22.3 (2.7) | 34.4 (NA) | 53.0 (NA) | 60.3 (7.3) | 45.6 (22.8) | ||||||
| 7− (3+/−2+/−) | 66.9 (NA) | 76.9 (NA) | 58.2 (16.6) | 68.3 (6.6) | 45.3 (NA) | 36.1 (NA) | 19.4 (9.5) | 43.0 (33.8) | ||||||
| Samples (n) | 1 | 1 | 3 | 2 | 1 | 1 | 3 | 2 | ||||||
| 3−5+2− | 2.2 (NA) | 10.8 (NA) | 3.3 (2.2) | 14.6 (20.0) | 3.3 (NA) | 6.4 (NA) | 6.1 (4.4) | 17.6 (22.1) | ||||||
| 3−5+2+ | 0.2 (NA) | 0.2 (NA) | 1.3 (1.1) | 0.5 (0.6) | 0.3 (NA) | 0.3 (NA) | 1.0 (0.6) | 1.2 (0.8) | ||||||
| 3+5+2− | 5.8 (NA) | 7.9 (NA) | 2.8 (0.7) | 2.4 (1.5) | 9.1 (NA) | 11.1 (NA) | 6.5 (0.8) | 2.9 (2.1) | ||||||
| 3+5+2+ | 12.8 (NA) | 14.7 (NA) | 22.7 (12.8) | 24.1 (8.0) | 32.7 (NA) | 52.2 (NA) | 60.7 (9.0) | 49.6 (24.5) | ||||||
| 5− (3+/−2+/−) | 79.0 (NA) | 66.5 (NA) | 69.9 (14.8) | 58.4 (30.1) | 54.5 (NA) | 30.0 (NA) | 25.6 (9.0) | 28.7 (0.6) | ||||||
| All Values Mean % (SD) . | IELs . | . | . | . | LPLs . | . | . | . | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Age . | Small intestine . | . | Large intestine . | . | Small intestine . | . | Large intestine . | . | ||||||
| . | <9 mo . | >9 mo . | <9 mo . | >9 mo . | <9 mo . | >9 mo . | <9 mo . | >9 mo . | ||||||
| Samples (n) | 6 | 3 | 13 | 8 | 6 | 3 | 13 | 8 | ||||||
| % Lymphocytes CD3+ Mean (SD) | 11.4 (10.5) | 28.0 (3.1) | 21.0 (15.7) | 33.8 (13.6) | 28.03 (18.0) | 64.27 (0.5) | 51.59 (27.0) | 54.18 (18.1) | ||||||
| Samples (n) | 4 | 1 | 7 | 4 | 4 | 1 | 7 | 4 | ||||||
| 3+4+ | 18.2 (24.1) | 46.9 (NA) | 20.2 (16.5) | 41.4 (24.5) | 52.5 (14.9) | 57.7 (NA) | 57.6 (11.9) | 51.5 (9.9) | ||||||
| 3+8+ | 20.2 (15.5) | 26.9 (NA) | 34 (23.1) | 31.3 (10.6) | 19.9 (8.4) | 24.3 (NA) | 24.6 (6.7) | 27.7 (9.3) | ||||||
| 3+4+8+ | 3.3 (2.3) | 3 (NA) | 1.9 (1.8) | 3.5 (4.2) | 4.7 (3.9) | 1.7 (NA) | 2.2 (2.1) | 6.4 (5.2) | ||||||
| 3+4−8− | 58.3 (36.8) | 23.2 (NA) | 43.9 (33.9) | 23.9 (15.1) | 22.9 (14.0) | 16.3 (NA) | 15.6 (10.0) | 14.5 (3.8) | ||||||
| Samples (n) | 1 | 1 | 3 | 2 | 1 | 1 | 3 | 2 | ||||||
| 3−7+2− | 8.7 (NA) | 0.5 (NA) | 9.3 (7.6) | 3.9 (5.2) | 5.9 (NA) | 0.8 (NA) | 5.7 (3.6) | 1.9 (2.3) | ||||||
| 3−7+2+ | 5.3 (NA) | 0.5 (NA) | 6.2 (3.2) | 3.4 (3.4) | 6.2 (NA) | 1.2 (NA) | 8.2 (4.4) | 6.0 (6.9) | ||||||
| 3+7+2− | 5.6 (NA) | 6.5 (NA) | 2.5 (0.5) | 2.0 (0.7) | 8.2 (NA) | 8.9 (NA) | 6.4 (0.8) | 3.5 (1.8) | ||||||
| 3+7−2+ | 13.5 (NA) | 15.7 (NA) | 23.7 (14.2) | 22.3 (2.7) | 34.4 (NA) | 53.0 (NA) | 60.3 (7.3) | 45.6 (22.8) | ||||||
| 7− (3+/−2+/−) | 66.9 (NA) | 76.9 (NA) | 58.2 (16.6) | 68.3 (6.6) | 45.3 (NA) | 36.1 (NA) | 19.4 (9.5) | 43.0 (33.8) | ||||||
| Samples (n) | 1 | 1 | 3 | 2 | 1 | 1 | 3 | 2 | ||||||
| 3−5+2− | 2.2 (NA) | 10.8 (NA) | 3.3 (2.2) | 14.6 (20.0) | 3.3 (NA) | 6.4 (NA) | 6.1 (4.4) | 17.6 (22.1) | ||||||
| 3−5+2+ | 0.2 (NA) | 0.2 (NA) | 1.3 (1.1) | 0.5 (0.6) | 0.3 (NA) | 0.3 (NA) | 1.0 (0.6) | 1.2 (0.8) | ||||||
| 3+5+2− | 5.8 (NA) | 7.9 (NA) | 2.8 (0.7) | 2.4 (1.5) | 9.1 (NA) | 11.1 (NA) | 6.5 (0.8) | 2.9 (2.1) | ||||||
| 3+5+2+ | 12.8 (NA) | 14.7 (NA) | 22.7 (12.8) | 24.1 (8.0) | 32.7 (NA) | 52.2 (NA) | 60.7 (9.0) | 49.6 (24.5) | ||||||
| 5− (3+/−2+/−) | 79.0 (NA) | 66.5 (NA) | 69.9 (14.8) | 58.4 (30.1) | 54.5 (NA) | 30.0 (NA) | 25.6 (9.0) | 28.7 (0.6) | ||||||
The phenotype of IEL and LPL fractions of cells derived from the small and the large intestine were analyzed by FACS. Cells were gated as lymphocytes based on known boundaries of forward scatter (FSC) and side scatter (SSC) defined with PBLs and LPLs, then gated as CD3− and CD3+ lymphocytes based on CD3 expression. Isotype negative controls were used to determine the CD3+/− threshold, and the position of the quadrants within the CD3+ and CD3− populations. CD3low cells were included with the CD3+ population for analysis of CD4/CD8 expression. For the purpose of CD2/CD5 and CD2/CD7 pre-T cell marker analysis, cells expressing low levels of CD3 but above the nonspecific threshold (determined with IgG isotype controls) were included as CD3− cells as these were most likely to contain immature pre-T cells. Results of CD3/CD4/CD8 expression are expressed as a percentage of the total lymphocytes satisfying both the CD3+ gate and the FSC/SSC lymphocyte gate, mean percentage values (SD). Results of CD2/CD7 and CD2/CD5 expression are expressed as a percentage of total lymphocytes within the FSC/SSC lymphocyte gate, mean percentage values (SD). The proportion of DN cells decreased with age, while that of total SP cells (CD3+4+ and CD3+8+) increased, and that of DP cells changed little. In analysis of infants aged <5 years, CD3+CD4−CD8− (DN) cells were negatively correlated with age: IEL −0.86 and −0.64 (for ileum and colon, respectively) and LPL −0.41 and −0.79. Pre-T cells expressing CD7 (CD3−CD7+2+/− by FACS) tended to decrease with age (NS).