Figure 1.
Representative gel picture showing restriction digestion of FOXP3 −3279 (rs 3761548) (a) and −2383 (rs 3761549) (b) for genotyping
(a) L1-L3: Heterozygous (487,329,158bp), L4 and L5: Homozygous wild (329,158bp), L6 and L7: Homozygous variant (487bp), M=100bp.
(b) L1, L2, L4, L9, L10: Homozygous (184,127,77 bp), L3, L5, L6, L7: Heterozygous (261,184,127,77 bp), L8: Homozygous variant (261, 127bp), M= 100bp.
bp=base pair, M=marker.
Table 1.
Demographic and clinical characteristics of study RPL and Control participants.
| Clinico-pathological characteristics | Cases Mean ± SD | Control Mean ± SD | Chi Square Test p value |
|---|---|---|---|
| Age | 29.83 ± 4.47 | 29.58 ± 3.98 | 0.690 |
| Live Birth | 0.40 ± 0.66 | 2.38 ± 0.62 | <0.0001 |
| Number of Pregnancies | 3.35 ± 1.14 | 2.65 ± 0.82 | <0.0003 |
| Miscarriages | 2.81 ± 1.08 | 0.00 ± 0.00 | - |
| Women with no live birth | 44 (36%) | 0 | - |
| Family History | |||
| YES | 23 (19.1%) | 15 (10%) | Ref |
| NO | 97 (80.8%) | 135 (90%) | 0.03 |
| Consanguinity | |||
| YES | 38 (31.6%) | 27 (18%) | Ref |
| NO | 82 (68.4%) | 123 (82%) | 0.01 |
Table 2.
Genotypic and allelic frequencies of FOXP3 3279 C/A and 2383 C/T polymorphism in RPL cases and controls.
| SNP | Genotype/Allele | Cases (%) N=120 | Controls (%) N=150 | OR (95% C.I) | p value |
|---|---|---|---|---|---|
| FOXp3-3279 C/A | CC | 30 (25) | 69 (46) | Ref | Ref |
| AC | 76 (63.3) | 69 (46) | 2.53 (1.47–4.34) | 0.0006 | |
| AA | 14 (11.7) | 12 (8) | 2.68 (1.1–6.4) | 0.03 | |
| AC+AA | 90 (75) | 81 (54) | 2.5 (1.5–4.3) | 0.0005 | |
| Alleles | C | 136 (56.6) | 207 (69) | Ref | Ref |
| A | 104 (43.3) | 93 (31) | 1.70 (1.1–2.4) | 0.003 | |
| FOXp3-2383 C/T | CC | 68 (56.6) | 95 (63.4) | Ref | Ref |
| CT | 42 (35) | 51 (34) | 1.15 (0.6–1.9) | 0.60 | |
| TT | 10 (8.3) | 04 (2.6) | 3.49 (1.0–11.6) | 0.04 | |
| CT+TT | 52 (43.3) | 55 (36.6) | 1.32 (0.8–2.1) | 0.31 | |
| Alleles | C | 178 (74.2) | 241 (80.3) | Ref | Ref |
| T | 62 (25.8) | 59 (19.7) | 1.42 (0.9–2.1) | 0.09 |
Table 3.
Combined genotypes of FOXP3 −3279 C/A and −2383 C/T and risk of RPL with respect to clinical characteristics.
| Marker | Parameter | Cases n=120 | Controls (N=150) | O.R (95%CI) | Fishers Exact Test p value | |||
|---|---|---|---|---|---|---|---|---|
| CC | AC+AA | CC | AC+AA | |||||
| rs3761548 | Age | ≤30 years | 20 (16.6%) | 54 (45%) | 39 (26%) | 33 (22%) | 3.1 (1.5–6.3) | 0.001 |
| >30 years | 10 (8.3%) | 36 (30%) | 30 (20%) | 48 (32%) | 2.2 (0.9–5.1) | 0.07 | ||
| F/H | Yes | 5 (4.1%) | 18 (15%) | 5 (3.3%) | 10 (6.6%) | 1.8 (0.4–7.7) | 0.47 | |
| No | 25 (20.8) | 72 (60%) | 64 (42.6%) | 71 (47.3%) | 2.5 (1.4–4.5) | 0.0009 | ||
| Cons. | CM | 9 (7.5%) | 29 (24.1%) | 6 (4%) | 21 (14%) | 0.9 (0.2–3.0) | 1 | |
| NCM | 21 (17.5%) | 61 (50.8%) | 63 (42%) | 60 (40%) | 3.0 (1.6–5.6) | 0.0002 | ||
| rs3761549 | CC | CC+TT | CC | CC+ TT | ||||
| Age | ≤30 years | 40 (33.3%) | 34 (28.3%) | 51 (34%) | 21 (14%) | 2.0 (1.0–4.0) | 0.04 | |
| >30 years | 28 (23.3%) | 18 (15%) | 44 (29.3%) | 34 (22.6%) | 0.8 (0.3–1.7) | 0.7 | ||
| F/H | Yes | 11 (9.16%) | 12 (10%) | 8 (5.3%) | 7 (4.6%) | 1.2 (0.3–4.5) | 0.9 | |
| No | 57 (47.5%) | 40 (33.3%) | 87 (58%) | 48 (32%) | 1.2 (0.7–2.1) | 0.4 | ||
| Cons. | CM | 18 (15%) | 20 (16.6%) | 17 (11.3%) | 10 (6.6%) | 1.8 (0.6–5.1) | 0.3 | |
| NCM | 50 (41.6%) | 32 (26.6%) | 78 (52%) | 45 (30%) | 1.1 (0.6–1.9) | 0.7 | ||
Table 4.
Results of analysis for the selected SNPs with RPL risk and other diseases.
| S. No. | Study | Country/Ethnicity | Case/Control | Disease | p value |
|---|---|---|---|---|---|
| 3279 FOXP3 rs3761548 | |||||
| 1. | Naderi et al., 2015 [13] | Iran/Asian | 195/101 | RPL | 0.387 |
| 2. | Saxena et al., 2015 [14] | India/Asian | 200/300 | RPL | <0.001 |
| 3. | Jabar et al., 2014 [19] | Palestine/Asian | 100/100 | RPL | 0.005 |
| 4. | Wu et al., 2012 [20] | China/Asian | 146/112 | RPL | 0.003 |
| 5. | Sharif et al., 2016 [21] | Palestine/Asian | 100/100 | RPL | 0.011 |
| 6. | Dirsipam et al., 2021 [22] | India/Asian | 150/150 | RPL | <0.001 |
| 7. | Hadinedoshan et al., 2015 [24] | Iran/Asian | 80/80 | RPL | 0.30 |
| 8. | Mishra et al., 2018 [25] | India/Asian | 100/100 | RPL | 0.18 |
| 9. | Our study (2024) | India/Asian | 120/150 | RPL | <0.001 |
| 2383 Foxp3 rs3761549 | |||||
| 1. | Song et al., 2012 [27] | China/Asian | 408/363 | Psoriasis vulgaris | <0.05 |
| 2. | Lan et al., 2010 [28] | China/Asian | 120/160 | SLE | <0.05 |
| 3. | Andre et al., 2011 [29] | Brazil/SA | 177/171 | Endometriosis | 0.003 |
| 4. | Tian et al., 2018 [30] | China/Asian | 560/582 | Breast Cancer | n. s |
| 5. | Our study (2024) | India/Asian | 120/150 | RPL | 0.04 |