Possible Role of Regulatory B Cells in Different Behçet’s Disease Phenotypes and Therapies: First Report from Egypt

Figures & data

Figure 1 Flow cytometric detection of regulatory B cells.

Notes: (A) Forward and side scatter histogram was used to define the lymphocytes population (R1). (B) The CD19+ cells (R2) were assessed within the lymphocyte population. (C) The expression of CD24 and CD38 was assessed in CD19+ cells to define CD19+CD24^+highCD38^+high cells (regulatory B cells).

Table 1 Clinical and Immunological Characteristics of the Study Subjects

	BD Patients N=35 N (%)	Control N=39	p-value
Arterial disease	17 (48.6%)	NA	NA
DVT	16 (45.7%)	NA	NA
GIT	10 (28.6%)	NA	NA
Skin lesions/Pathergy test positive/Oral ulcers	35 (100%)	NA	NA
Genital ulcers	16 (45.7%)	NA	NA
Urethritis, n (%)	6 (17.1%)	NA	NA
Ocular disease		NA	NA
• Anterior uveitis	2 (5.7%)
• Pan uveitis and retinal vasculitis	6 (17.1%)
• Pan uveitis and retinal vasculitis with optic neuritis/atrophy	12 (34.3%)
• No ocular signs	15 (42.9%)
Arthralgia/arthritis	20 (57.1%)	NA	NA
Headache	5 (14.3%)	NA	NA
Seizures	0	NA	NA
Cognitive dysfunction	6 (17.1%)	NA	NA
Peripheral neuropathy	10 (28.6%)	NA	NA
Psychosis	0	NA	NA
Inflammatory back pain	14 (40%)	NA	NA
Activity within the past four weeks (BDCAF 2006)	21 (60%)	NA	NA
Erectile dysfunction (IIEF-5 score) #	15 (9–20)	20 (20–25)	<0.001^*
Depression score (Beck questionnaire) #	13 (8–19)	10 (7–14)	0.042^*
BSAS #	30 (30–60)	NA	NA
ESR#	25 (17–34)	20 (16–26)	0.02^*
CRP§	14.4 ± 9.8	2 ± 3.4	<0.001^*
B lymphocytes % §	11.1 ±1.9	14.2 ± 3	<0.001^*
B reg.% §	1.9 ± 0.5	3.3 ± 1.4	<0.001^*

Notes: Independent T test, Mann-Whitney U-test and Chi-square test were used as appropriated.

* Significance was considered when P<0.05

Abbreviations: BD, Behcetʹs disease; SD, standard deviation; DVT deep venous thrombosis; GIT gastrointestinal manifestations; BDCAF, Behçetʹs Disease Current Activity Form; #, median (interquartile range); §, mean± standard deviation; BSAS, Behçetʹs Syndrome Activity Score; ESR, erythrocyte sedimentation rate; CRP, C-reactive protein.

Table 2 DMARDs Administration in BD Patients

Drug Used	BD Patients N=35, N (%)
Prednisolone doses
10 mg	9 (25.7%)
20 mg	11 (31.4%)
30 mg	7 (20%)
40 mg	7 (20%)
60 mg	1 (2.9%)
Prednisolone duration (years) #	4 (3–5)
CYC	8 (22.9%)
CS	11 (31.4%)
MTX	9 (25.7%)
AZA	13 (37.1%)
LDA	12 (34.3%)
Anticoagulant	4 (11.4%)

Abbreviations: DMARDS, disease-modifying anti-rheumatic drugs; BD, Behçetʹs disease; #, median (interquartile range); CYC, cyclophosphamide; CS, cyclosporine; MTX, methotrexate; AZA, azathioprine; LDA, low dose aspirin.

Table 3 The Differences in Total B Lymphocytes and B Regulatory Subset with Different Disease Characteristics

	% of B Cells in (+) Group Median (IQR)	% of B Cells in (-) Group Median (IQR)	p-value	% of B Reg. in (+) Group Median (IQR)	% of B Reg. in (-) Group Median (IQR)	p-value
Activity within 4 weeks	12 (11–12.4)	11.3 (10.4–12)	0.14	1.86 (I.5–2.3)	I.8 (I.5–2.2)	0.5
Arterial disease	11 (9–12)	12 (11.3–12.5)	0.013^*	2.2 (I.7–2.4)	I 7 (I.5–2)	0.067
DVT	11.8 (10.7–12.1)	11.5 (10.4–12.4)	0.8	2.1 (I.8–2.4)	I.6 (I.4–2.I)	0.052
GIT	12 (11.3–12.4)	11.4 (10.4–12.2)	0.3	1.9 (I.7–2.6)	I.8 (I.5–2.2)	0.4
Genital ulcers	11.6 (10.2–12.1)	11.9 (10.8–12.4)	0.4	2.1 (I.7–2.5)	I.7 (I.4-I.9)	0.04^*
Urethritis	11.8 (11.2–12.2)	11.6 (10.4–12.2)	0.7	2.5 (2.2–2.6)	I.8 (I.5–2)	0.002^*
Arthralgia/arthritis	11.9 (10.5–12.6)	11.4 (10.4–12.1)	0.46	I.8 (I.4–2.4)	I.86 (I.6–2.I)	0.88
Headache	12.2 (11.2–12.4)	11.6 (10.4–12.2)	0.57	2 (I.6–2.I6)	I.8 (I.5–2.2)	0.84
PN	11.3 (7.2–12)	12 (10.6–12.4)	0.12	I.9 (I.5–2.3)	I.86 (I.5–2.2)	0.76
Cognitive dysfunction	12.2 (11.2–12.4)	11.5 (10.4–12)	0.38	2 (I.6–2.I6)	I.8 (I.5–2.2)	0.69
Inflammatory back pain	11.6 (10.6–12.2)	11.9 (10.4–12.2)	0.93	I.8 (I.5–2.2)	I.86 (I.6–2.2)	0.82
Smoking	10.6 (9.4–12.2)	11.9 (10.8–12.2)	0.32	2.3 (I.5–2.5)	I.8 (I.5–2.I)	0.24
Family history of AIDs	12 (10.4–12.3)	11.5 (10.4–12.2)	0.92	2 (I.7–2.4)	I.8 (I.4–2.2)	0.3

Note: Mann-Whitney U-test was used to identify the significance between the groups.

* Significance was considered when P<0.05.

Abbreviations: IQR, interquartile range; DVT, deep venous thrombosis; GIT, gastrointestinal manifestations; PN, peripheral neuropathy; AID, autoimmune diseases.

Figure 2 The relationship of B lymphocytes with ESR and BSAS scores.

Abbreviations: ESR, erythrocyte sedimentation rate; BSAS, Behçet’s Syndrome Activity Score.

Figure 3 The differences in medians of total B lymphocytes and B regs. Percentages with versus without different drug administration. *Significant difference.

Abbreviations: CYC, cyclophosphamide; CSA, cyclosporine-A; MTX, methotrexate; AZA, azathioprine Anticoag; anticoagulant; LDA, low dose aspirin.