Differentiation of regulatory B cells in a mouse model of plasma cell mastitis

doi:10.3877/cma.j.issn.1674-0807.2025.05.003

Abstract

Abstract:

Objective

To investigate the regulatory B cell (Breg) differentiation in a mouse model of plasma cell mastitis （PCM）.

Methods

Fifteen female BALB/c mice were randomly divided into three groups： PCM model group receiving 80 μl intramammary injection of oil in water emulsion containing pathological tissue homogenate from one PCM patient，Freund's adjuvant group receiving complete Freund's adjuvant injection of the same amount into the breast and control group receiving physiological saline injection of the same amount into the breast. HE staining was used to observe the pathological manifestations of mouse breast tissue. Enzyme linked immunosorbent assay （ELISA） was used to detect the expression of inflammatory factors in mouse serum，flow cytometry was used to detect the proportion of Breg in spleen，and immunofluorescence was used to detect the expression of CD21 and CD138 in mouse breast tissue. Continuous data were compared among multiple groups using analysis of variance，and pairwise comparisons were conducted using the LSD test.

Results

The HE staining results showed plasma cell infiltration in the breast tissue of mice in the PCM model group. The ELISA results showed that the expressions of inflammatory factors IL-10，TNF-α，IL-6，IL-35 and IL-1β in the PCM model group mice were significantly higher than those in the control group and Freund's adjuvant group （all P<0.001）. The flow cytometry results showed that the proportion of CD19⁺CD5^-CD1d⁺IL10⁺ Breg in the PCM model group was significantly higher than that in the control group and Freund's adjuvant group （24.66% vs 3.09%，P<0.001； 24.66% vs 2.53%，P<0.001），and the proportion of CD19⁺CD5⁺CD1d⁺IL10⁺ Breg was significantly increased compared with the control group and Freund's adjuvant group （11.66% vs 2.92%，P<0.001； 11.66% vs 1.93%，P<0.001）. The proportion of CD21 positive cells in the breast tissue of mice in the PCM model group was significantly increased compared with the control group and Freund's adjuvant group （36.45% vs 16.77%，P<0.001； 36.45% vs 16.47%，P<0.001），and the proportion of CD138 positive cells was significantly increased compared with the control group and Freund's adjuvant group （17.18% vs 7.33%，P<0.001； 17.18% vs 7.48%，P<0.001）.

Conclusion

The PCM mouse model shows abnormal differentiation of Breg.

Key words: Plasma cell mastitis, Inflammatory factors, Immune cells, Regulatory B cells

Chi Pan, Wei Jiang, Hongyang Kong, Qingtao Ni. Differentiation of regulatory B cells in a mouse model of plasma cell mastitis[J]. Chinese Journal of Breast Disease(Electronic Edition), 2025, 19(05): 275-281.

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Figures/Tables 7

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组别	只数	IL-10	TNF-α	IL-4	IL-6	IL-35	IL-1β
对照组	5	286.40±34.77	259.46±61.95	82.44±24.14	82.03±15.73	16.42±6.55	64.67±13.55
弗氏佐剂组	5	280.41±42.38	268.55±115.39	86.91±17.81	80.62±14.52	17.31±6.33	68.95±25.95
PCM模型组	5	603.01±37.94^ab	856.10±110.89^ab	97.45±23.17	201.86±17.36^ab	38.88±7.93^ab	219.83±26.69^ab
F值		114.945	59.534	0.620	95.598	16.647	74.656
P值		<0.001	<0.001	0.554	<0.001	<0.001	<0.001

组别	只数	IL-10	TNF-α	IL-4	IL-6	IL-35	IL-1β
对照组	5	286.40±34.77	259.46±61.95	82.44±24.14	82.03±15.73	16.42±6.55	64.67±13.55
弗氏佐剂组	5	280.41±42.38	268.55±115.39	86.91±17.81	80.62±14.52	17.31±6.33	68.95±25.95
PCM模型组	5	603.01±37.94^ab	856.10±110.89^ab	97.45±23.17	201.86±17.36^ab	38.88±7.93^ab	219.83±26.69^ab
F值		114.945	59.534	0.620	95.598	16.647	74.656
P值		<0.001	<0.001	0.554	<0.001	<0.001	<0.001

组别	只数	CD19⁺CD5^-CD1⁺IL10⁺	CD19⁺CD5⁺CD1⁺IL10⁺
对照组	5	3.09±0.55	2.92±1.57
弗氏佐剂组	5	2.53±1.16	1.93±1.24
PCM模型组	5	24.66±3.15^ab	11.66±2.32^ab
F值		124.213	27.382
P值		<0.001	<0.001

组别	只数	CD19⁺CD5^-CD1⁺IL10⁺	CD19⁺CD5⁺CD1⁺IL10⁺
对照组	5	3.09±0.55	2.92±1.57
弗氏佐剂组	5	2.53±1.16	1.93±1.24
PCM模型组	5	24.66±3.15^ab	11.66±2.32^ab
F值		124.213	27.382
P值		<0.001	<0.001

组别	只数	CD21	CD138
对照组	5	16.77±4.19	7.33±3.19
弗氏佐剂组	5	16.47±4.46	7.48±2.02
PCM模型组	5	36.45±7.54^ab	17.18±2.44^ab
F值		20.884	5.951
P值		<0.001	0.016

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