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Different Intraepithelial CD3(+) Cell Numbers in Crohns Disease and Ulcerative Colitis

Celia Escudero-Hernández, Enrique Montalvillo, Beatriz Antolin, David Bernardo, Jose Antonio Garrote, Eduardo Arranz and Luis Fernandez-Salazar

The self-archived postprint version of this journal article is available at Linköping University Institutional Repository (DiVA):

http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-164197

N.B.: When citing this work, cite the original publication.

Escudero-Hernández, C., Montalvillo, E., Antolin, B., Bernardo, D., Antonio Garrote, J., Arranz, E., Fernandez-Salazar, L., (2020), Different Intraepithelial CD3(+) Cell Numbers in Crohns Disease and Ulcerative Colitis, Inflammatory Bowel Diseases, 26(3), E14-E15. https://doi.org/10.1093/ibd/izz309

Original publication available at:

https://doi.org/10.1093/ibd/izz309

Copyright: Oxford University Press (OUP) (Policy B - Oxford Open Option B - CC-BY)

http://www.oxfordjournals.org/

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Page 1 of 8

Title: Different intraepithelial CD3

+

cell numbers in Crohn’s disease and ulcerative 1

colitis 2

Authors: Celia Escudero-Hernández, PhD

1,*

, Enrique Montalvillo, PhD

1

, Beatriz 3

Antolín, MD

2

, David Bernardo, PhD

1

, José Antonio Garrote, MD, PhD

1,3

, Eduardo 4

Arranz, MD, PhD

1

, Luis Fernández-Salazar, MD, PhD

2

5

Affiliations:

6

1

Mucosal Immunology Laboratory, Instituto de Biología y Genética Molecular (IBGM), 7

University of Valladolid-CSIC, C/ Sanz y Forés 3, 47003 Valladolid, Spain.

8

2

Digestive Disease Unit. Hospital Clínico Universitario de Valladolid. Avda Ramón y 9

Cajal 3, 47003 ,Valladolid, Spain.

10

3

Laboratory of Molecular Genetics, Hospital Universitario Rio Hortega, C/ Dulzaina 2, 11

47012 Valladolid, Spain.

12

*Current address: Department of Clinical and Experimental Medicine (IKE), Linköping 13

University, Linköping, Sweden 14

Corresponding authors:

15

Celia Escudero-Hernández. Department of Clinical and Experimental Medicine 16

(IKE), Ingång 68, Plan 13, US campus, Linköping University, s-581 83, Linköping, 17

Sweden. E-mail: celia.escher@gmail.com / celia.escudero-hernandez@liu.se 18

 Eduardo Arranz. Mucosal Immunology Laboratory, Instituto de Biología y 19

Genética Molecular (IBGM), University of Valladolid-CSIC, C/ Sanz y Forés 3, 20

47003 Valladolid, Spain. Tel.No: +34 983 18 42 31. Fax: +34 983 18 48 12. E- 21

mail: earranz@uva.es 22

Acknowledgements: This work has been partially funded by Junta de Castilla y León 23

(SAN/673/2008 and grant to E.M.) and University of Valladolid (grant to C.E.H.).

24

Competing interests: The authors declare that they have no conflict of interest.

25

Keywords: Diagnosis, flow cytometry, IBD, mucosal immunology.

26

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Page 2 of 8

Summary: Intraepithelial immune response can be studied by flow cytometry. The 27

proportion of CD3+ cells differs between Crohn’s disease and ulcerative colitis.

28

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Page 3 of 8 To the editors,

29

We read with interest the paper by Roosenboom et al. entitled “Intestinal CD103

+

CD4

+

30

and CD103

+

CD8

+

T-cell subsets in the gut of inflammatory bowel disease patients at 31

diagnosis and during follow-up”.

1

The authors found a decreased number of mucosal 32

CD103

+

CD4

+

and CD103

+

CD8

+

T-cells in both inflammatory bowel disease (IBD) types, 33

Crohn’s disease (CD) and ulcerative colitis (UC), which recovered after clinical 34

remission. However, CD103

-

CD4

+

T-cells were more abundant in both CD and UC. With 35

a similar technical approach, we aimed at characterizing the overall numbers of T-cells 36

(CD45

+

CD3

+

) in a Spanish CD and UC patient cohort, although focusing on the 37

intraepithelial mucosal layer. We enrolled a total of 13 CD (8 colonic and 9 ileal samples) 38

and 14 UC (colonic samples) patients, all of them endoscopically active. CD and UC 39

were diagnosed using current guidelines.

2 3

In addition, 8 non-IBD patients with inflamed 40

mucosa (6 colonic and 4 ileal samples), and 15 patients with no macroscopic or 41

microscopic findings (11 colonic and 7 ileal samples) were included as inflamed (IC) and 42

healthy (Hc) controls, respectively (table 1). Detailed patient diagnosis data and 43

materials and methods can be found in the supplementary data file.

44

The proportion of intraepithelial CD45

+

cells was lower in the colon of inflamed controls 45

compared to healthy controls (figure 1A). However, this population remained unchanged 46

in CD (in both colon and ileum) and UC (figure 1A-B) regarding to healthy controls, which 47

is in agreement with previous evidence.

4

On the other side, the proportion ofT-cells in 48

UC was increased, which resulted in a decreased number of CD45

+

CD3

-

cells. Thus, the 49

T/CD3

-

ratio was higher in UC patients compared to CD and the healthy control groups 50

(figure 1C). CD ileal T/CD3- ratio, as in the colon, remained normal (figure 1D). These 51

results suggests that the intraepithelial compartment in active UC might behave different 52

from active CD. Therefore, the T-cell subsets should be studied both in the intraepithelial 53

layer and the lamina propria compartments separately. We are aware that our cohort is 54

small and further research is needed. Still, the difference observed in T-cell and CD3

-

55

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Page 4 of 8

numbers in the intraepithelial compartment might be a useful tool for differential 56

diagnosis of UC and it would be worth to explore in depth in UC and other forms of colitis.

57

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Page 5 of 8 References

58

1. Roosenboom B, Wahab PJ, Smids C, et al. Intestinal CD103+CD4+ and 59

CD103+CD8+ T-Cell Subsets in the Gut of Inflammatory Bowel Disease Patients 60

at Diagnosis and During Follow-up. Inflamm Bowel Dis 2019;25(9):1497-509.

61

2. Gomollon F, Dignass A, Annese V, et al. 3rd European Evidence-based Consensus 62

on the Diagnosis and Management of Crohn's Disease 2016: Part 1: Diagnosis 63

and Medical Management. J Crohns Colitis 2017;11(1):3-25.

64

3. Magro F, Gionchetti P, Eliakim R, et al. Third European Evidence-based Consensus 65

on Diagnosis and Management of Ulcerative Colitis. Part 1: Definitions, 66

Diagnosis, Extra-intestinal Manifestations, Pregnancy, Cancer Surveillance, 67

Surgery, and Ileo-anal Pouch Disorders. J Crohns Colitis 2017;11(6):649-70.

68

4. Smids C, Horjus Talabur Horje CS, Drylewicz J, et al. Intestinal T Cell Profiling in 69

Inflammatory Bowel Disease: Linking T Cell Subsets to Disease Activity and 70

Disease Course. J Crohns Colitis 2018;12(4):465-75.

71

72

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Page 6 of 8 TABLES

73

Table 1 Clinical and demographic characteristics of IBD patients and controls

Variable Hc IC CD UC

Total number of subjects

15 8 13 14

No. of colonic / ileal samples

11 / 7 6 / 4 8 / 9 14 / 0

Gender, % females 60.00% 62.50% 26.67% 43.75%

Median age (range) 52 (31-71) 51 (26-83) 27 (17-48) 46 (26-79) Median CRP levels

(range)

N/A N/A 17 (0-31) 3 (0-284)

Montreal UC extent,

%

N/A N/A N/A

E1: ulcerative proctitis

3 (21.43%)

E2: left-sided UC 4 (28.57%)

E3: extensive UC 7 (50.00%)

Montreal CD location, n (%)

N/A N/A N/A

L1: ileal 6 (46.16%)

L2: colonic 3 (23.08%)

L3: ilealcolonic 4 (30.76%)

Behaviour N/A N/A N/A

B1: nonstructuring, nonpenetrating

10 (76.92%)

B2: structuring 3 (23.08%)

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Page 7 of 8

CD, Crohn’s disease; Hc, healthy controls; IBD, inflammatory bowel disease; IC, inflamed non-IBD controls; N/A, not applicable; UC, ulcerative colitis.

74

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Page 8 of 8 Figure legends

75

Figure 1: Intraepithelial T-cells are increased in ulcerative colitis. (A-B) Percentage 76

of intraepithelial CD45

+

cells in colonic (A) and ileal (B) samples. (C-D) T-cell/CD3

-

cell 77

ratio in intraepithelial colonic (C) and ileal (D) samples. Biopsy samples were obtained 78

from healthy controls (Hc), non-IBD inflamed controls (IC), Crohn’s disease (CD), and 79

ulcerative colitis (UC) patients. Boxes represent data within the interquartile range and 80

the horizontal bar represents the population median. Statistically significant differences 81

are shown as * p<0.05.

82

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Figure 1

A B C D

16 *

12

8

0

+% CD45 colonic cells 4

Hc IC CD UC

*

* 16

12

8

0 4 Ratio T/CD3- colonic cells

Hc IC CD UC 16

12

8

0

% CD45+ ileal cells 4

Hc IC CD

16

12

8

0

-Ratio T/CD3 ileal cells 4

Hc IC CD

References

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