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Flow cytometric analysis of the CD4+ TCR Vβ repertoire in the peripheral blood of children with type 1 diabetes mellitus, systemic lupus erythematosus and age-matched healthy controls.

Tzifi F, Kanariou M, Tzanoudaki M, Mihas C, Paschali E, Chrousos G, Kanaka-Gantenbein C - BMC Immunol. (2013)

Bottom Line: Data regarding the quantitative expression of TCR Vβ subpopulations in children with autoimmune diseases provided interesting and sometimes conflicting results.Statistical analysis revealed that the CD4 + Vβ4 chain was significantly increased in patients with T1DM (p < 0.001), whereas CD4 + Vβ16 one was significantly increased in SLE patients (p < 0.001) compared to controls.CD4 + Vβ4 and CD4 + Vβ16 chains could be possibly involved in the cascade of events precipitating the pathogenesis of T1DM and SLE in children, respectively.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: Data regarding the quantitative expression of TCR Vβ subpopulations in children with autoimmune diseases provided interesting and sometimes conflicting results. The aim of the present study was to assess by comparative flow cytometric analysis the peripheral blood CD4+ TCR Vβ repertoire of children with an organ-specific autoimmune disorder, such as type 1 diabetes mellitus (T1DM), in comparison to children with a systemic autoimmune disease, such as Systemic Lupus Erythematosus (SLE) in comparison to healthy age-matched controls of the same ethnic origin. The CD4+ TCR Vβ repertoire was analysed by flow cytometry in three groups of participants: a) fifteen newly diagnosed children with T1DM (mean age: 9.2 ± 4.78 years old), b) nine newly diagnosed children with SLE, positive for ANA and anti-dsDNA, prior to treatment (mean age: 12.8 ±1.76 years old) and c) 31 healthy age-matched controls (mean age: 6.58 ± 3.65 years old), all of Hellenic origin.

Results: CD4 + TCR Vβ abnormalities (± 3SD of controls) were observed mainly in SLE patients. Statistical analysis revealed that the CD4 + Vβ4 chain was significantly increased in patients with T1DM (p < 0.001), whereas CD4 + Vβ16 one was significantly increased in SLE patients (p < 0.001) compared to controls.

Conclusions: CD4 + Vβ4 and CD4 + Vβ16 chains could be possibly involved in the cascade of events precipitating the pathogenesis of T1DM and SLE in children, respectively.

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Comparison of CD4 + Vβ4 chain values among the 3 groups of children studied. Values (%) of the CD4 + Vβ4 chain are higher in the majority of T1DM children in comparison to controls.
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Figure 4: Comparison of CD4 + Vβ4 chain values among the 3 groups of children studied. Values (%) of the CD4 + Vβ4 chain are higher in the majority of T1DM children in comparison to controls.

Mentions: The increased/decreased usage of a Vβ subfamily was defined as control mean value ± 2 or 3 standard deviations [10,11]. Five patients (33%) presented no abnormalities in all CD4 + Vβ chains values when compared to controls (Table 3, Figure 2). One T1DM patient had decreased usage of Vβ14 (0.7%, mean value: 4.00 ± 1.78) and in another one Vβ20 value was 0.1% (mean value: 4.08 ± 1.75). None of the healthy individuals had such low usage of these Vβ chains. Increased usage of several CD4 + Vβ chains (Vβ3, Vβ4, Vβ5.1, Vβ5.3, Vβ8, Vβ11, Vβ18, Vβ16 and Vβ21.3) was identified sporadically in T1DM patients. With the exception of Vβ11 chain, increased usage of the mentioned Vβ subpopulations were also found in healthy individuals. Although the CD4 + Vβ4 lymphocyte population increased usage was found in 5 T1DM patients when compared to the mean value of controls, statistical analysis showed a significant difference between the two groups (p < 0.001) concerning the CD4 + Vβ4 subfamily (Table 4, Figure 3). This is expected, since values of the controls ranged between 0.0% and 2.1% while T1DM values ranged from 0.6% to 3.3%. It should be mentioned that 17 healthy children had values of the CD4 + Vβ4 chain < 1% and 12 T1DM children had values > 1% (Figure 4). Taking into account the very low number of CD4 + Vβ4 cells analyzed and that the maximum coefficient of variation (CV) was 20%, statistical analysis was adjusted to this CV. As shown in Table 5, the differences remained significant even after “narrowing” the mean values, adjusting for a 20% difference in the mean values of our measurements.


Flow cytometric analysis of the CD4+ TCR Vβ repertoire in the peripheral blood of children with type 1 diabetes mellitus, systemic lupus erythematosus and age-matched healthy controls.

Tzifi F, Kanariou M, Tzanoudaki M, Mihas C, Paschali E, Chrousos G, Kanaka-Gantenbein C - BMC Immunol. (2013)

Comparison of CD4 + Vβ4 chain values among the 3 groups of children studied. Values (%) of the CD4 + Vβ4 chain are higher in the majority of T1DM children in comparison to controls.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3750582&req=5

Figure 4: Comparison of CD4 + Vβ4 chain values among the 3 groups of children studied. Values (%) of the CD4 + Vβ4 chain are higher in the majority of T1DM children in comparison to controls.
Mentions: The increased/decreased usage of a Vβ subfamily was defined as control mean value ± 2 or 3 standard deviations [10,11]. Five patients (33%) presented no abnormalities in all CD4 + Vβ chains values when compared to controls (Table 3, Figure 2). One T1DM patient had decreased usage of Vβ14 (0.7%, mean value: 4.00 ± 1.78) and in another one Vβ20 value was 0.1% (mean value: 4.08 ± 1.75). None of the healthy individuals had such low usage of these Vβ chains. Increased usage of several CD4 + Vβ chains (Vβ3, Vβ4, Vβ5.1, Vβ5.3, Vβ8, Vβ11, Vβ18, Vβ16 and Vβ21.3) was identified sporadically in T1DM patients. With the exception of Vβ11 chain, increased usage of the mentioned Vβ subpopulations were also found in healthy individuals. Although the CD4 + Vβ4 lymphocyte population increased usage was found in 5 T1DM patients when compared to the mean value of controls, statistical analysis showed a significant difference between the two groups (p < 0.001) concerning the CD4 + Vβ4 subfamily (Table 4, Figure 3). This is expected, since values of the controls ranged between 0.0% and 2.1% while T1DM values ranged from 0.6% to 3.3%. It should be mentioned that 17 healthy children had values of the CD4 + Vβ4 chain < 1% and 12 T1DM children had values > 1% (Figure 4). Taking into account the very low number of CD4 + Vβ4 cells analyzed and that the maximum coefficient of variation (CV) was 20%, statistical analysis was adjusted to this CV. As shown in Table 5, the differences remained significant even after “narrowing” the mean values, adjusting for a 20% difference in the mean values of our measurements.

Bottom Line: Data regarding the quantitative expression of TCR Vβ subpopulations in children with autoimmune diseases provided interesting and sometimes conflicting results.Statistical analysis revealed that the CD4 + Vβ4 chain was significantly increased in patients with T1DM (p < 0.001), whereas CD4 + Vβ16 one was significantly increased in SLE patients (p < 0.001) compared to controls.CD4 + Vβ4 and CD4 + Vβ16 chains could be possibly involved in the cascade of events precipitating the pathogenesis of T1DM and SLE in children, respectively.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: Data regarding the quantitative expression of TCR Vβ subpopulations in children with autoimmune diseases provided interesting and sometimes conflicting results. The aim of the present study was to assess by comparative flow cytometric analysis the peripheral blood CD4+ TCR Vβ repertoire of children with an organ-specific autoimmune disorder, such as type 1 diabetes mellitus (T1DM), in comparison to children with a systemic autoimmune disease, such as Systemic Lupus Erythematosus (SLE) in comparison to healthy age-matched controls of the same ethnic origin. The CD4+ TCR Vβ repertoire was analysed by flow cytometry in three groups of participants: a) fifteen newly diagnosed children with T1DM (mean age: 9.2 ± 4.78 years old), b) nine newly diagnosed children with SLE, positive for ANA and anti-dsDNA, prior to treatment (mean age: 12.8 ±1.76 years old) and c) 31 healthy age-matched controls (mean age: 6.58 ± 3.65 years old), all of Hellenic origin.

Results: CD4 + TCR Vβ abnormalities (± 3SD of controls) were observed mainly in SLE patients. Statistical analysis revealed that the CD4 + Vβ4 chain was significantly increased in patients with T1DM (p < 0.001), whereas CD4 + Vβ16 one was significantly increased in SLE patients (p < 0.001) compared to controls.

Conclusions: CD4 + Vβ4 and CD4 + Vβ16 chains could be possibly involved in the cascade of events precipitating the pathogenesis of T1DM and SLE in children, respectively.

Show MeSH
Related in: MedlinePlus