Detection of human cytomegalovirus in synovial neutrophils obtained from patients with rheumatoid arthritis

Figures & data

Table 1. Results from polymerase chain reaction (PCR), direct immunofluorescence (IF), and serology analyses of samples from rheumatoid arthritis (RA) patients, and healthy controls

Peripheral blood (%)								Synovial fluid (%)
		Positive	Negative	No RNAse P	No sample	Uncertain	Σ	Positive	Negative	No RNAse P	No sample	Uncertain	Σ	Σ^δ
Rheumatoid arthritis
PCR	HCMV-IE DNA	24 (73)	9	1*	–	0	34	13 (41)	19	2*	–	0	34	30 (88)
	HCMV-pp150 DNA	16 (67)	8	1*	9*	0	34	13 (57)	10	3*	8*	–	34
IF	HCMV-IEA	0	16	–	–	1	17	0	17	–	–	–	17	13 (76)
	HCMV-pp65	3 (18)	12	–	–	2	17	11 (65)	3	–	–	3	17
ELISA	HCMV-IgM	1 (3)	27	–	–	5	33	N/A	N/A	–	–	–		1 (3)
	HCMV-IgG	25 (76)	8	–	–	–	33	N/A	N/A	–	–	–		25 (76)
Healthy controls
ELISA	HCMV IgM	0	71	–	–	–	71	–	–	–	–	–	–	0
	HCMV IgG	54 (76)	17	–	–	–	71	–	–	–	–	–	–	54 (76)

ELISA, enzyme-linked immunosorbent assay; IE, immediate early; IEA, immediate early antigen; IgG, immunoglobulin G; IgM, immunoglobulin M; N/A, not available; Σ, total number of samples, Σ^δ, total HCMV-positive samples. *Excluded from data analysis.

Figure 1. (A) Human cytomegalovirus (HCMV) pp65 in polymorphonuclear leucocytes (PMNLs) (ethanol fixed) from synovial fluid (SF) obtained from the knee of a patient with rheumatoid arthritis (RA). (B) HCMV-pp65 in in-vitro HCMV-infected PMNLs obtained from a healthy donor. (C) Distribution of positive samples for immediate early (IE) and pp150 DNA in PMNLs from peripheral blood (PB) and SF, respectively, in patients with RA. Total number of analysed samples was, for PB IE, n = 33; pp150, n = 24; SF IE, n = 32; SF pp150, n = 23. (D) Delta Ct values (mean and range, calculated by subtracting mean Ct of housekeeping gene from mean Ct of target gene) in samples positive for HCMV IE and/or pp150 DNA (mean Ct ≤ 40, at least one out of two or three reactions positive), in PB and SF, for HCMV IE DNA and pp150 DNA, respectively. PB IE, n = 24; SF IE, n = 13; PB pp150, n = 16; SF pp150, n = 13

Table 2. Results from polymerase chain reaction, enzyme-linked immunosorbent assay, and direct immunofluorescence analyses of polymorphonuclear leucocytes from peripheral blood (PB) and synovial fluid (SF) of rheumatoid arthritis patients, including data on anti-citrullinated protein antibody (ACPA) positivity, gender, and age at sampling

PB		SF
IE-DNA	pp150-DNA	IE-DNA	pp150-DNA	HCMV-IgM	HCMV-IgG	ACPA	IF PB IEA	IF PB pp65	IF SF IE	IF SF pp65	Gender	Age at sampling (years)
Positive	Positive	Negative	Positive	Equivocal	Positive	Positive	Negative	Negative	Negative	Positive	F	35
Positive	No sample	Positive	No sample	Equivocal	Positive	Positive	Negative	Negative	Negative	Positive	F	80
Positive	No sample	Negative	Negative	Negative	Positive	Positive	Negative	Negative	Negative	Positive	F	57
Positive	No sample	Negative	Positive	Equivocal	Positive	Positive	Negative	Positive	Negative	Positive	F	39
Negative	No sample	Negative	No sample	Negative	Positive	Positive	Negative	Negative	Negative	Positive	F	67
Positive	Positive	Positive	Positive	Negative	Positive	Positive	Negative	Negative	Negative	Positive	F	59
Negative	Negative	Positive	No sample	Negative	Negative	Positive	Negative	Negative	Negative	Positive	F	37
Positive	Negative	Negative	Negative	Negative	Negative	Positive	Negative	Negative	Negative	Positive	M	29
Positive	Positive	Positive	Negative	Negative	Negative	Positive	?	Negative	Negative	Positive	M	38
Positive	No sample	Negative	No RNAse P	Negative	Positive	Positive	Negative	Negative	Negative	Positive	F	59
Negative	Positive	Positive	Negative	Negative	Negative	Positive	Negative	Negative	Negative	Positive	F	59
Negative	Positive	Positive	Positive	Negative	Positive	Positive	Negative	Positive	Negative	Negative	F	80
Negative	No sample	Negative	No sample	Equivocal	Positive	Positive	Negative	Negative	Negative	Negative	M	60
Negative	Negative	Positive	Positive	Equivocal	Positive	Positive	Negative	Negative	Negative	Negative	F	81
Positive	Negative	Positive	Positive	Negative	Positive	Positive	Negative	Positive	Negative	Negative	M	50
Positive	Negative	Negative	Negative	Negative	Positive	Positive	Negative	?	Negative	?	F	50
Positive	No sample	Negative	No sample	Negative	Positive	Positive	Negative	?	Negative	?	F	41
Positive	No sample	Negative	No sample	Negative	Positive	Positive					M	39
Positive	Negative	Negative	No sample	Positive	Positive	Negative					F	34
Positive	Negative	Negative	Positive			Positive					F	73
No RNAse P	No RNAse P	Negative	No sample	Negative	Positive	Positive					F	64
Negative	Positive	Negative	Negative	Negative	Positive	?					F	35
Negative	No sample	Negative	Negative	Negative	Positive	?					F	44
Negative	Positive	No RNAse P	No RNAse P	Negative	Positive	Positive					F	69
Positive	Positive	Positive	Positive	Negative	Negative	Positive					F	33
Positive	Positive	Negative	Negative	Negative	Positive	Positive					M	59
Positive	Positive	Positive	Positive	Negative	Positive	Positive					F	47
Positive	Positive	Negative	Positive	Negative	Positive	Positive					F	35
Positive	Positive	Positive	Positive	Negative	Positive	Positive					F	71
Positive	Positive	Negative	Negative	Negative	Positive	Positive					F	68
Positive	Positive	Positive	Positive	Negative	Negative	Positive					F	82
Positive	Positive	No RNAse P	No RNAse P	Negative	Negative	Positive					F	37
Positive	Negative	Positive	Positive	Negative	Negative	Positive					F	62
Positive	Positive	Negative	Negative	Negative	Positive	Positive					M	30

HCMV, human cytomegalovirus; IE, immediate early; IF, immunofluorescence; IgG, immunoglobulin G; IgM, immunoglobulin M. Question marks denote equivocal results.