Immunity Networks in Early Detection and Diagnosis of Lung Cancer

Oleg Kshivets, MD, PhDDepartment of Surgery, Siauliai Cancer Center

LithuaniaThe 4th International Congress on Lung Cancer

Halkidiki, Greece, 2001

The Main Characteristics of The Study Populations

• Lung Cancer Patients (males – 472, females – 61; age=57.40.4 years)………………………..……..533

• Stage I………………………………………………...48• Stage II………………………………………………..47• Stage III………………………………………………321• Stage IV………………………………………………117• Lung Non-Malignant Pathology (males – 188,

females – 94; age=50.30.7 years) ……………..282• Non-Malignant Tumor..……………………………100• Abscess………………………..…………………….112• Tuberculoma…………………..……………………..70

Procedures (n=815)• Lung Cancer

Patients……………533• Pneumonectomy…181• Upper Lobectomy..138• Lower Lobectomy....67• Upper/Lower

Bilobectomy…….….24• Middle Lobectomy.…6• Segmentectomy.….76• Exploratory Thoraco-

tomy, biopsy……….41

• Lung Non-Malignant Pathology…………282

• Pneumonectomy…...5• Upper Lobectomy…96• Lower

Lobectomy....81• Middle Lobectomy.…2• Segmentectomy,

Wedge Resection….98

Lung Cancer Patients Characteristics (n=533)

• Squamous Cell..…294• Adenocarcinoma..171• Large Cell………….48• Small Cell………….20• T1.…………………116• T2………………….168• T3………………….125• T4………………….124• N0………………….148

• N1………………….144• N2………………….159• N3……………………82• G1…………………...88• G2………………….166• G3………………….279• M0………………….438• M1…………………...95

Factors of T-, B-, K-cell Circuit: Lymphocytes (%); Lymphocytes (abs)*109/l; Lymphocytes (tot) *109 T-cells (%); T-cells (abs)*109/l; T-cells (tot)*109

B-cells (%); B-cells (abs)*109/l; B-cells (tot)*109 K-cells (%); K-cells (abs)*109/l; K-cells (tot)*109 CD1 (%); CD1 (abs)*109/l; CD1 (tot)*109 CDw26 (%); CDw26 (abs)*109/l; CDw26 (tot)*109 CD4+2H (%); CD4+2H (abs)*109/l; CD4+2H (tot) *109 CD8+VV (%); CD8+VV (abs)*109/l; CD8+VV (tot) *109 CD4 (%); CD4 (abs)*109/l; CD4 (tot) *109 CD8 (%); CD8 (abs)*109/l; CD8 (tot) *109 CD4/CD8 Index Thymus Function Factors of Monocyte and Macrophage Circuit: Monocytes (%); Monocytes (abs)*109/l; Monocytes (tot) *109 Phagocyte Index; Phagocyte Number Index Complete Phagocytosis Factors of Humoral Immunity: IgG (g/l); IgM (g/l); IgA (g/l) Natural Antibodies Circulating Immune Complexes Factors of Neutrophil Circuit: NST spontaneous NST stimulated Staphylococcus aureus NST stimulated Streptococcus Pyogenes Index Stimulation by Staphylococcus aureus Index Stimulation by Streptococcus Pyogenes Leukocytes (abs)*109/l; Leukocytes (tot)*109 Stick Neutrophils (%); Stick Neutrophils (abs)*109/l Stick Neutrophils (tot)*109 Segmented Neutrophils (%); Segmented Neutrophils (abs)*109/l; Segmented Neutrophils (tot)*109

Immune Testing

Table 1. Factors of T-, B- and K-cell Circuit between LCP

(n=95) and NMP (n=282) in terms of Early Detection NN Factors LCP st I-II n=95 NMP n=282 P Mean SD Mean SD 1 2 3

B-cells (%) B-cells (abs)*109/l B-cells (tot)*109

18.34 0.39 1.94

8.32 0.44 2.21

16.46 0.29 1.44

6.46 0.14 0.76

0.024* 0.001* 0.001*

4 5 6

Monocytes (%) Monocytes (abs)*109/l Monocytes (tot) *109

3.45 0.22 1.08

2.92 0.22 1.10

2.87 0.16 0.76

2.29 0.14 0.70

0.048* 0.003* 0.001*

7 IgM (g/l) 1.56 0.51 1.72 0.62 0.021* 8 Index Stimulation by

Streptococcus pyogenes 1.37

0.94

1.89

2.13

0.024*

9 Segmented Neutrophils (tot)*109

17.92

8.46

16.06

7.28

0.040*

Table 2. Factors of T-, B- and K-cell Circuit between LCP and NMP in terms of Diagnosis

NN Factors LCP st I-IV n=533 NMP n=282 P Mean SD Mean SD 1 B-cells (%) 17.89 7.74 16.46 6.46 0.009* 2 3

CD4+2H (abs)*109/l CD4+2H (tot) *109

0.53 2.56

0.36 1.82

0.59 2.87

0.38 1.94

0.034* 0.027*

4 CD4 (tot) *109 3.41 2.25 3.78 2.38 0.030* 5 6

CD8 (abs)*109/l CD8 (tot) *109

0.22 1.05

0.17 0.81

0.25 1.22

0.20 0.98

0.015* 0.012*

7 Lymphocytes (%) 28.04 11.12 33.07 11.43 0.000* 8 9 10

Monocytes (%) Monocytes (abs)*109/l Monocytes (tot) *109

3.24 0.21 1.01

2.57 0.21 1.06

2.87 0.16 0.76

2.29 0.14 0.70

0.045* 0.000* 0.000*

11 12

IgM (g/l) IgA (g/l)

1.61 2.52

0.58 1.02

1.72 2.26

0.62 0.85

0.012* 0.000*

13 14 15 16 17 18 19 20

Leukocytes (abs)*109/l Leukocytes (tot)*109 Stick Neutrophils (%) Stick Neutrophils (abs)*109/l Stick Neutrophils (tot)*109 Segmented Neutrophils (%) Segmented Neutrophils (abs)*109/l Segmented Neutrophils (tot)*109

6.31 30.09 1.27 0.09 0.44 64.23 4.10 19.49

2.48 12.52 1.57 0.18 0.78 11.48 1.88 9.06

5.47 26.73 0.97 0.06 0.28 59.47 3.29 16.06

1.85 10.19 1.58 0.12 0.57 11.75 1.41 7.28

0.000* 0.000* 0.010* 0.003* 0.003* 0.000* 0.000* 0.000*

Table 3. Results of Logistic Regression Modeling in Early Detection of Lung Cancer (n=95) from NMP (n=282)

NN Significant Immune Factors: 2=27.543; Df=9; P=0.0011407; Estimate 1 2 3 4 5 6 7 8 9

Const.B0 B-cells (%) B-cells (abs)*109/l B-cells (tot)*109

Monocytes (%) Monocytes (abs)*109/l Monocytes (tot) *109 IgM (g/l) Index Stimulation by Streptococcus pyogenes Segmented Neutrophils (tot)*109

1.34429 -0.01543 -3.80134 0.45341 0.00167 2.36224

-0.70182 0.44811 0.18113

-0.01816

ExpectedNormal

Frequency Distribution: Residuals (LCP I-II, n=95-----NMP, n=282)Logistic Regression Modeling in Early Detection of Lung Cancer

Chi2=27.543; df=9; P=0.0011; n=377

No

of o

bs

0102030405060708090

100110120130

-1.1-1.0

-0.9-0.8

-0.7-0.6

-0.5-0.4

-0.3-0.2

-0.10.0

0.10.2

0.30.4

0.50.6

0.70.8

Table 4. Results of Logistic Regression Modeling in Diagnosis of LC (n=533) from NMP (n=282)

NN Significant Immune Factors: 2=108.075; Df=20; P=0.0000000; Estimate 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20

Const.B0 B-cells (%) CD4+2H (abs)*109/l CD4+2H (tot) *109

CD4 (tot) *109 CD8 (abs)*109/l CD8 (tot) *109 Lymphocytes (%) Monocytes (%) Monocytes (abs)*109/l Monocytes (tot) *109 IgM (g/l) IgA (g/l) Leukocytes (abs)*109/l Leukocytes (tot)*109 Stick Neutrophils (%) Stick Neutrophils (abs)*109/l Stick Neutrophils (tot)*109 Segmented Neutrophils (%) Segmented Neutrophils (abs)*109/l Segmented Neutrophils (tot)*109

4.67194 -0.02946 3.17501

-0.57679 0.09542 3.60816

-0.41372 -0.03025 0.07468

-0.39635 -0.53516 0.43660

-0.37333 -1.40877 0.22663

-0.08793 3.97775

-0.64340 -0.05288 1.09895

-0.18262

ExpectedNormal

Frequency Distribution: Residuals (LCP I-IV, n=533-----NMP, n=282)Logistic Regression Modeling in Diagnosis of Lung Cancer

Chi2=108.075; df=20; P=0.0000000; n=815

No

of o

bs

020406080

100120140160180200220240260

-1.2 -1.0 -0.8 -0.6 -0.4 -0.2 0.0 0.2 0.4 0.6 0.8 1.0 1.2

Results of Multi-Factor Clustering in Early Detection of Lung Cancer

Results of Multi-Factor Clustering in Diagnosis of Lung Cancer

SEPATH-models in Early Detection of Lung Cancer

Early Diagnosis of Lung CancerNormalized Residuals (SEPATH-model of Immune Cell Circuit)

LCP stage I-II (n=95)---NMP (n=282)Chi2=38749.1; k=989; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-4

-3

-2

-1

0

1

2

3

4

-15 -10 -5 0 5 10 15 20 25

Early Detection of Lung CancerNormalized Residuals (SEPATH-model of Macrophage Circuit)

LCP stage I-II (n=95)---NMP (n=282)Chi2=662.8; k=20; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-2.5

-1.5

-0.5

0.5

1.5

2.5

-10 -8 -6 -4 -2 0 2 4 6 8

Early Detection of Lung CancerNormalized Residuals (SEPATH-model of Humoral Immunity)

LCP stage I-II (n=95)---NMP (n=282) Chi2=585.9; k=9; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-2.5

-1.5

-0.5

0.5

1.5

2.5

-10 -8 -6 -4 -2 0 2 4 6 8

Early Detection of Lung CancerNormalized Residuals (SEPATH-model of Neutrophils Circuit)

LCP stage I-II (n=95)---NMP (n=282) Chi2=5214.4; k=77; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-3

-2

-1

0

1

2

3

-12 -8 -4 0 4 8 12 16

SEPATH-models in Diagnosis of Lung Cancer

Diagnosis of Lung CancerNormalized Residuals (SEPATH-model od Humoral Immunity)

LCP stage I-IV (n=533)---NMP (n=282)Chi2=243.1; k=9; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-2.5

-1.5

-0.5

0.5

1.5

2.5

-14 -12 -10 -8 -6 -4 -2 0 2 4

Diagnosis of Lung CancerNormalized Residuals (SEPATH-model of Neutrophils Circuit)

LCP stage I-IV (n=533)---NMP (n=282) Chi2=10772.3; k=77; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-3

-2

-1

0

1

2

3

-15 -10 -5 0 5 10 15 20 25

Diagnosis of Lung CancerNormalized Residuals (SEPATH-model of Immune Cell Circuit)

LCP stage I-IV (n=533)-----NMP (n=282)Chi2=80569.9; k=989; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-4

-3

-2

-1

0

1

2

3

4

-25 -15 -5 5 15 25 35

Diagnosis of Lung CancerNormalized Residuals (SEPATH-model of Macrophage Circuit)

LCP stage I-IV (n=533)---NMP (n=282)Chi2=312.1; k=20; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-2.5

-1.5

-0.5

0.5

1.5

2.5

-14 -10 -6 -2 2 6 10

Results of Monte Carlo Simulation in Early

Detection and Diagnosis of Lung Cancer

Diagnosis of Lung Cancer (LCP st I-IV, n=533---NMP, n=282)Normalized Residuals

SEPATH-Model: Diagnosis-Cell Circuit-Humoral Immunity- Monocyte/Macrophage Circuit-Neutrophils Circuit

Chi2=62865.7; k=1426; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-4

-3

-2

-1

0

1

2

3

4

-30 -20 -10 0 10 20 30

0.927 1.055 1.182 1.309 1.436 1.564 1.691 1.818 1.945 2.073 above

Monte Carlo Data: Diagnosis of Lung Cancer (LCP st I-IV, n=533---NMP, n=282) Diagnosis-Cell Circuit-Humoral Immunity-Monocyte/Macrophage Circuit-Neutrophils

(Bootstrap) Chi2=10330.03; k=1426; P=0.000000

Early Detection of Lung Cancer (LCP st I-II, n=95---NMP, n=282)Normalized Residuals

SEPATH-Model: Early Detection-Cell Circuit-Humoral Immunity- Monocyte/Macrophage Circuit-Neutrophils Circuit

Chi2=31850.5; k=1426; P=0.000000

Value

Exp

ecte

d N

orm

al V

alue

-4

-3

-2

-1

0

1

2

3

4

-25 -15 -5 5 15 25

0.901 1.002 1.103 1.203 1.304 1.405 1.506 1.607 1.708 1.809 above

Monte Carlo Data: Early Detection of Lung Cancer (LCP st I-II,n=95---NMP,n=282)Early Detection-Cell Circuit-Hum. Immunity-Mon./Macrophage Circuit-Neutrophils

(Bootstrap) Chi2=11089.4; k=1421; P=0.000000

T-, B-Cell Circuit in Early Detection of Lung Cancer

T-, B-Cell Circuit in Diagnosis of Lung Cancer

CD8, CD4 Cell Circuit in Diagnosis of Lung Cancer

Lymphocytes, Monocytes Circuit in Early Detection of Lung Cancer

Lymphocytes, Monocytes Circuit in Diagnosis of Lung Cancer

Neutrophils Circuit in Early Detection of Lung Cancer

Neutrophils Circuit in Diagnosis of Lung Cancer

Humoral Immunity in Early Detection of Lung Cancer

Humoral Immunity in Diagnosis of Lung Cancer

0 2 4 6 8 10 122

0

2

4

6Model "Early Cancer---Immune System"

Lung Cancer Cell Population

Imm

une

Cyt

otox

ic C

ell P

opul

atio

ns

5.072

2.072

X1 3

X2 3

X3 3

X4 3

10.0590.235 X1 2 X2 2

X3 2 X4 2

0 5 10 15 200

5

10Model "Early Cancer---Immune System"

Time

Cel

l Pop

ulat

ion

Dyn

amic

s

5.884

0.306

X1 2

X1 3

200 X1 1

0 5 10 15 20 25 3010

5

0

5

10Model "Invasive Cancer---Immune System"

Lung Cancer Cell Population

Imm

une

Cyt

otox

ic C

ell P

opul

atio

ns

10.187

8.334

X1 3

X2 3

X3 3

X4 3

25.4662.403 10 14 X1 2 X2 2

X3 2 X4 2

0 5 100

10

20

30Model "Invasive Cancer---Immune System"

Time

Cel

l Pop

ulat

ion

Dyn

amic

s

25.07

8.691 10 14

X4 2

X4 3

100 X4 1

Lotka-Volterra Models of Lung Cancer Cell Population and Cytotoxic Cell Population Dynamics

Holling-Tenner Models of Lung Cancer Cell Population and Cytotoxic Cell Population Dynamics

0 5 100

2

4

6Model "Early Cancer---Immune System"

Lung Cancer Cell Population

Imm

une

Cyt

otox

ic C

ell P

opul

atio

ns

4.266

0.414

X1 3

X2 3

X3 3

X4 3

100.103 X1 2 X2 2

X3 2 X4 2

0 50 100 150 200 2500

2

4

6Model "Early Cancer---Immune System"

Time

Cel

l Pop

ulat

ion

Dyn

amic

s

4.266

0.103

X1 2

X1 3

2000 X1 1

0 5 10 15 200

5

10

15Model "Invasive Cancer---Immune System"

Lung Cancer Cell Population

Imm

une

Cyt

otox

ic C

ell P

opul

atio

ns

13.007

1

X1 3

X2 3

X3 3

X4 3

19.5124 X1 2 X2 2

X3 2 X4 2

0 20 40 600

10

20Model "Invasive Cancer---Immune System"

Time

Cel

l Pop

ulat

ion

Dyn

amic

s

19.51

1

X4 2

X4 3

500 X4 1

Immunity Networks in Early Detection of Lung Cancer

Immunity Networks in Diagnosis of Lung Cancer

Lung Cancer Dynamics

Networks between Immune System and Recognition of Lung Cancer

SUPERONCOIMMUNOLOGY-1.0

IMMUNODIAGNOSIS-1 IMMUNODIAGNOSIS-2

IMMUNODEFICIENCY NORMMALIGNANT NEOPLASM

POPULATION PHASE TRANSITION IMMUNOSTAGING

N M G

EARLY CANCER INVASIVE CANCER

Conclusions:• early detection and diagnosis of lung

cancer significantly depended on:• 1) the level of immune cell circuit;• 2) the value of humoral immunity

factors;• 3) the level of monocyte and

macrophage circuit;• 4) the value of neutrophils circuit.

Address:

• Oleg Kshivets, M.D., Ph.D.• Thoracic Surgeon• Department of Surgery• Siauliai Cancer Center• Tilzes:42-16, 5400 Siauliai, Lithuania• Tel. (37041)416614• Fax (37041)526430• okshivets@yahoo.com

kshivets@hotmail.com• http//:myprofile.cos.com/Kshivets