Patterns of hepatitis b virus immune escape and pol/rt mutations across clinical cohorts of patients with genotypes a, e and occult hepatitis b infection in Nigeria: A multi-centre study

Figures & data

Figure 1. Distribution of study participants across clinical cohorts of HBV infection.

Table 1. Socio-demographic characteristics and HBV serological profiles of study participants.

		Gender		HBsAg		Anti-HBs		HBeAg		Anti-HBc IgM		Anti-HBc IgG
Clinical Cohorts	Frequency (%)	Male (%)	Female (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)
Serological Profile of Clinical Cohorts of Patients
Acute HBV Infection	5 (5.3)	5 (100.0)	0 (0.0)	5 (100.0)	0 (0.0)	0 (0.0)	5 (100.0)	1 (20.0)	4 (80.0)	5 (100.0)	0 (0.0)	5 (100.0)	0 (0.0)
Chronic HBV Infection	59 (62.1)	29 (49.2)	30 (50.8)	59 (100.0)	0 (0.0)	1 (1.7)	58 (98.3)	13 (22.0)	46 (78.0)	2 (3.4)	57 (96.6)	55 (93.2)	4 (6.8)
Chronic HBV with LC	6 (6.3)	6 (100.0)	0 (0.0)	6 (100.0)	0 (0.0)	0 (0.0)	6 (100.0)	1 (16.7)	5 (83.3)	0 (0.0)	6 (100.0)	6 (100.0)	0 (0.0)
Chronic HBV with HCC	18 (18.9)	15 (83.3)	3 (16.7)	18 (100.0)	0 (0.0)	0 (0.0)	18 (100.0)	7 (38.9)	11 (61.1)	12 (66.7)	6 (33.3)	18 (100.0)	0 (0.0)
Occult HBV Infection	7 (7.4)	2 (28.6)	5 (71.4)	0 (0.0)	7 (100.0)	0 (0.0)	7 (100.0)	1 (14.3)	6 (85.7)	0 (0.0)	7 (100.0)	7 (100.0)	0 (0.0)
Total	95 (100.0)	57 (60.0)	38 (40.0)	88 (92.6)	7 (7.4)	1 (1.1)	94 (98.9)	23 (24.2)	72 (75.8)	19 (20.0)	76 (80.0)	91 (95.8)	4 (4.2)
		Gender		HBsAg		Anti-HBs		Anti-HBc IgM		Anti-HBc IgG		HBeAg
Age group (yrs)	Frequency N(%)	Male (%)	Female (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)	Pos (%)	Neg (%)
Serological Profile of Total Study Population
21–30	29 (30.5)	12 (41.4)	17 (58.6)	28 (96.6)	1 (3.4)	1 (3.4)	28 (96.6)	3 (10.3)	26 (89.7)	27 (93.1)	2 (6.9)	6 (20.7)	23 (79.3)
31–40	29 (30.5)	21 (72.4)	8 (27.6)	27 (93.1)	2 (6.9)	0 (0.0)	29 (100.0)	4 (13.8)	25 (86.2)	27 (93.1)	2 (6.9)	8 (27.6)	21 (72.4)
41–50	20 (21.1)	14 (70.0)	6 (30.0)	19 (95.0)	1 (5.0)	0 (0.0)	20 (100.0)	5 (25.0)	15 (75.0)	20 (100.0)	0 (0.0)	4 (20.0)	16 (80.0)
51–60	15 (15.8)	10 (66.7)	5 (33.3)	12 (75.0)	3 (25.0)	0 (0.0)	15 (100.0)	6 (40.0)	9 (60.0)	15 (100.0)	0 (0.0)	5 (33.3)	10 (66.7)
61–70	2 (2.1)	0 (0.0)	2 (100.0)	2 (100.0)	0 (0.0)	0 (0.0)	2 (100.0)	1 (50.0)	1 (50.0)	2 (100.0)	0 (0.0)	0 (0.0)	2 (100.0)
Total	95 (100.0)	57 (60.0)	38 (40.0)	88 (92.6)	7 (7.4)	1 (1.1)	94 (98.9)	19 (20.1)	76 (79.9)	91 (95.8)	4 (4.2)	23 (24.2)	72 (75.8)

Note: LC – Liver Cirrhosis; HCC – Hepatocellular Carcinoma.

Pos: Positive for the test assay; Neg: Negative for the test assay.

Figure 2. Rectangular phylogenetic tree depicts the rare genotype A3 sequences obtained from this study. Tree Scale: 0.05 -

Figure 3. HBV S gene amino acid sequence alignment using bioedit software.

Table 2. Rate of HBV S gene pointmutation and mutation types across clinical cohorts of patients.

Clinical cohorts	Overall Sample, N	Cases with mutations, n (%)	Number of mutation types observed from all cases, N	Mutations	Mutation type rate in all cases, n (%)
Acute HBV	5	2 (40.0)	5	G159A K24R N59S S140L TS71	1 (25.0) 1 (25.0) 1 (25.0) 1 (25.0) 1 (25.0)
Chronic HBV	61	25 (40.9)	91	K24R R169A S113T Others	12 (12.63) 8 (8.42) 3 (3.16) 68 (71.58)
HBV with liver cirrhosis	5	2 (40.0)	7	R169A K24R M103K N59S S113T S117C/P	1 (12.5) 2 (25.0) 1 (12.5) 1 (12.5) 1 (12.5) 1 (12.5)
HBV with HCC	17	11 (64.7)	34	G44E K24R R169P S109P R169A Others	2 (5.9) 5 (14.7) 3 (8.8) 2 (5.9) 2 (5.9) 20 (58.8)
Occult HBV infection	7	5(71.4)	13	K24R S113T R169A Others	2 (12.5) 3 (18.8) 2 (12.5) 6 (37.5)

Note: HCC – Hepatocellular Carcinoma.

Others – All mutations asides those listed in each clinical cohort above.

Table 3. Socio-demographic, serological, and HBV S gene mutational profiles of enrollees with occult HBV infection.

Sample ID	Age (yrs)	Gender	Diagnosis	HBsAg	anti-HBsAb	HBeAg	anti-HBeAb	anti-HBc IgM	Anti-HBc IgG	HBV S gene Mutation
S114	56	Male	Hepatocellular Carcinoma	Negative	Negative	Negative	Negative	Negative	Positive	S113T+R169A
S143	50	Male	Hepatocellular Carcinoma	Negative	Negative	Negative	Negative	Negative	Positive	K24R
S150	43	Female	Apparently healthy subject	Negative	Negative	Positive	Negative	Negative	Positive	K24R, A45S, F134I,R169A,
S152	53	Female	Apparently healthy subject	Negative	Negative	Negative	Negative	Negative	Positive	K24R, G44E, A45S, L109P, G145A
S162	48	Female	Apparently healthy subject	Negative	Negative	Negative	Positive	Negative	Positive	-
S65	28	Female	Apparently healthy subject	Negative	Negative	Negative	Negative	Negative	Positive	S113T
S84	35	Female	Apparently healthy subject	Negative	Negative	Negative	Negative	Negative	Positive	S113T

Figure 4. HBV Pol/RT mutational analysis using bioedit software. image depicts HBV polymerase gene sequence at amino- acid position 115–223 with the amino-acid substitutions observed in the sequences.

Figure 5. Distribution of Pol gene RT domain amino-acid substitutions among 14 HBV isolates analysed.

Data Availability statement

The hepatitis B virus Polymerase and S gene sequences obtained from this study had been deposited in the Genbank sequence database under accession numbers OP420514 to OP420522, OP428653 to OP428701 https://mail.google.com/mail/u/0/#search/gb-admin%40ncbi.nlm.nih.gov/FMfcgzGqQSSScVjrPsPRrPvRSnpZmLjr.