Utility of Monocyte Expression of HLA-DR versus T Lymphocyte Frequency in the Assessment of COVID-19 Outcome

Figures & data

Figure 1 Gating strategies. Gating strategies for detecting the percentage of monocytes expressing HLA-DR and the mode of HLA-DR expression on monocytes and the frequency of T lymphocytes, with illustration of an example of mHLADR in mild to moderate case and severe case.

Table 1 Age, Demographic Data and the Clinical Disease Presentation of the Study Participants

		Group				P value
		Mild to Moderate Group (n=49)		Severe Group (n=48)
		Count	%	Count	%
Sex	M	21	42.9%	25	52.1%	0.363
	F	28	57.1%	23	47.9%
Fever	Yes	18	36.7%	39	81.3%	< 0.001
	No	31	63.3%	9	18.8%
Cough	Yes	20	40.8%	39	81.3%	< 0.001
	No	29	59.2%	9	18.8%
Dyspnea	Yes	14	28.6%	45	93.8%	< 0.001
	No	35	71.4%	3	6.3%
GIT manifestations	Yes	5	10.2%	6	12.5%	0.721
	No	44	89.8%	42	87.5%
CT Grade	0	2	4.1%	0	0.0%	< 0.001
	2	2	4.1%	0	0.0%
	3	37	75.5%	3	6.3%
	4	8	16.3%	28	58.3%
	5	0	0.0%	17	35.4%
DM	Yes	6	12.2%	17	35.4%	0.007
	No	43	87.8%	31	64.6%
Hypertension	Yes	10	20.4%	24	50.0%	0.002
	No	39	79.6%	24	50.0%
Bronchial asthma	Yes	8	16.3%	3	6.3%	0.118
	No	41	83.7%	45	93.8%
		Mean	SD	Mean	SD	P value
Age/year		42.22	14.51	60.31	14.48	< 0.001
O2 saturation %		96.02	1.39	80.75	11.4	< 0.001

Abbreviations: CT, chest computed tomography; DM, diabetes mellitus; GIT, gastro-intestinal tract; SD, standard deviation.

Table 2 Comparison Between Mild to Moderate Group and Severe Group Regarding Laboratory Data

	Mild to Moderate Group (n=49)			Severe Group (n=48)			P value
	Median	1st Quartile	3rd Quartile	Median	1st Quartile	3rd Quartile
ALT U/L	19.00	14.00	35.00	26.00	16.50	38.00	0.066
CREAT mg/dl	0.70	0.60	1.00	1.00	0.60	1.50	0.043
TLC (x10^3 \mm^3)	7.00	5.30	8.60	7.90	5.85	12.50	0.096
ANC (x10^3 \mm^3)	4.50	3.60	6.40	5.95	3.80	8.10	0.011
AMC (x10^3 \mm^3)	0.54	0.40	0.70	0.50	0.13	0.90	0.492
ALC (x10^3 \mm^3)	1.49	1.00	1.90	0.94	0.60	1.70	0.009
PLT (x10^3 \mm^3)	240.00	172.00	288.00	193.50	141.00	267.00	0.126
NLR	3.40	1.90	5.70	5.70	2.95	12.15	< 0.001
PLR	166.90	96.25	255.00	209.00	134.50	250.15	0.143
% of monocytes expressing mHLA-DR	98.00	97.00	99.00	80.50	53.00	93.00	< 0.001
mHLA-DR MFI	208.00	148.00	259.00	90.00	58.90	111.50	< 0.001
T lymphocytes	57.80	55.00	66.00	33.75	19.23	52.26	< 0.001
CRP mg/L	3.10	1.40	8.00	65.70	18.00	119.50	< 0.001
D-dimer ng/mL	400.00	300.00	450.00	900.00	445.00	2278.00	< 0.001

Abbreviations: ALC, absolute lymphocyte count; AMC, absolute monocytic count; ANC, absolute neutrophil count; CRP, C reactive protein; MLR, monocyte lymphocyte ratio; mHLA-DR, monocytes human leukocyte antigen D related; NLR, neutrophil lymphocyte ratio; PLR, platelet lymphocyte ratio; TLC, total leucocyte count.

Table 3 Comparison of Study Variables Between Survivors and Non-Survivors

	Outcome of Severe Cases						P value
	Severe Survivors (n=35)			Severe Non-Survivors (n=13)
	Median	Percentile 25	Percentile 75	Median	Percentile 25	Percentile 75
Monocytes % expressing HLADR	82.00	53.00	94.00	60.00	34.00	84.00	0.336
mHLADR MFI	93.00	61.00	113.00	64.00	40.00	83.00	0.070
T lymphocytes %	42.10	19.27	54.95	20.30	13.49	27.69	0.004
CRP mg/L	66.00	11.40	108.00	65.40	38.00	131.00	0.444
D-dimer ng/mL	900.00	400.00	1600.00	2101.00	730.00	3100.00	0.112

Abbreviations: CRP, C reactive protein; mHLA-DR, monocytes human leukocyte antigen D related.

Table 4 Correlation of mHLA-DR MFI, Percentage of Monocytes Expressing HLA-DR and T Lymphocyte Percentage with the Other Studied Markers in All Study Cases (n=97)

	mHLA-DR MFI		% of Monocytes Expressing MHLA-DR		T Lymphocyte %
	Correlation Coefficient	P value	Correlation Coefficient	P value	Correlation Coefficient	P value
O2 saturation %	0.632	< 0.001	−0.418	< 0.001	0.669	<0.001
PDW/fl	−0.050	0.625	0.187	0.067	−0.044	0.671
MPV /fl	0.258	0.011	−0.059	0.569	0.162	0.114
ALT U/L	−0.205	0.044	0.336	0.001	−0.328	0.001
Creatinine mg/dl	−0.091	0.374	−0.194	0.057	−0.216	0.034
TLC (x10^3 \mm^3)	−0.189	0.064	−0.158	0.122	−0.102	0.320
ANC (x10^3 \mm^3)	−0.257	0.011	−0.222	0.029	−0.151	0.139
AMC (x10^3 \mm^3)	0.113	0.271	−0.318	0.002	0.028	0.783
ALC (x10^3 \mm^3)	0.220	0.030	0.078	0.446	0.187	0.066
NLR	−0.309	0.002	0.279	0.006	−0.212	0.037
PLR	−0.071	0.491	−0.398	< 0.001	−0.173	0.090
T lymphocytes %	0.504	< 0.001	0.011	0.917	————	———–
CRP mg/L	−0.494	< 0.001	0.464	0.000	−0.536	<0.001
D-dimer ng/mL	−0.455	< 0.001	−0.477	0.000	−0.326	0.001

Abbreviations: ALC, absolute lymphocyte count; ALT, alanine transaminase; AMC, absolute monocytic count; ANC, absolute neutrophil count; CRP, C reactive protein; MLR, monocyte lymphocyte ratio; mHLA-DR, monocytes human leukocyte antigen D related; NLR, neutrophil lymphocyte ratio; PLR, platelet lymphocyte ratio; TLC, total leucocyte count.

Figure 2 Correlation of mHLA-DR MFI with other study markers in all study cases (n=97). (A) Positive correlation of mHLA-DR MFI with O2 saturation was seen (r=0.632, P < 0.001) (B) Negative correlation of mHLA-DR MFI with CRP was seen (r=−0.494, P < 0.001). (C) Positive correlation of mHLA-DR MFI with T lymphocyte % was seen (r=0.504, P < 0.001).

Table 5 (A) The ROC Curve Output Data for the Percentage of Monocytes Expressing HLA-DR, mHLA-DR MFI, T Lymphocytes Percentage, D-Dimer, and CRP as Discriminators of Severity Between Mild-to-Moderate and Severe Groups. (B) ROC Curve Output Data for T Lymphocyte Percentage Discrimination of Survival in Severe ICU Admitted Patients

	Area Under Curve		P value	95% Confidence Interval		Cut Off	Sensitivity %	Specificity %	PPV %	NPV %	Accuracy
				Lower Bound	Upper Bound
A: Output data of ROC curve for discrimination of severity.
mHLA-DR MFI	0.9		< 0.001	0.843	0.963	143	89.6	81.6	82.69	88.89	85.57
% of monocytes expressing HLA-DR	0.87		< 0.001	0.793	0.946	94.5	79	85.7	90.00	78.95	83.51
T lymphocytes %	0.9		< 0.001	0.839	0.960	54.4	81.3	81.6	94.44	77.05	83.51
CRP mg/L	0.87		< 0.001	0.796	0.948	15	77.1	85.7	97.06	76.19	83.51
D-dimer ng/mL	0.82		< 0.001	0.728	0.902	525	70.8	85.7	82.93	75.00	78.35
B: Output data of ROC curve for discrimination of survival.
T lymphocytes %	0.77		< 0.001	0.64	0.9	38.99	100	57	46.43	100.00	68.75

Abbreviations: CRP, C reactive protein; MFI, Mean fluorescence intensity; mHLA-DR, monocytes human leukocyte antigen D related; PPV, positive predictive value; NPV, negative predictive value.

Figure 3 Output data of ROC curves regarding monocyte % expressing HLA-DR, mHLA-DR MFI, T lymphocytes %. (A) Output data of ROC curve for monocyte % expressing HLA-DR, mHLA-DR MFI, T lymphocyte %, as discriminators of severity between mild to moderate group and severe group. (B) Output data of ROC curve to assess T lymphocyte frequency as a predictor of survival in ICU admitted patients (P< 0.001).

Table 6 Multiple Regression Analysis of Percentage of Monocytes Expressing HLADR, HLADR MFI, and T Lymphocyte Percentage with Co-Founders Affecting COVID-19 Severity and Survival

		P value	OR	95% C.I.
				Lower	Upper
Severity	Monocytes % expressing HLADR	0.307	0.946	0.850	1.052
	HLADR MFI	0.025	0.976	0.955	0.997
	T lymphocytes %	0.018	0.849	0.741	0.972
	Age	0.517	1.025	0.951	1.104
	DM	0.139	6.988	0.532	91.744
	Hypertension	0.836	0.782	0.076	8.064
	Bronchial asthma	0.161	0.035	0.000	3.787