Effect of neonatal thymectomy and antilymphocyte globulin on non-specific mitogen stimulation in Holstein-Friesian calves

Veterinary Immunology and Immunopathology, 2 (1981) 175--188 Elsevier Scientific Publishing C o m p a n y , A m s t e r d a m -- Printed in Belgium

175

EFFECT OF N E O N A T A L T H Y M E C T O M Y AND A N T I L Y M P H O C Y T E GLOBULIN ON N O N - S P E C I F I C MITOGEN S T I M U L A T I O N IN H O L S T E I N - F R I E S I A N CALVES

T.G.

SNIDER

III I

L.G.

ADAMS 2 and K.R.

PIERCE 2

IDepartment of V e t e r i n a r y Pathology, School of V e t e r i n a r y Medicine, L o u i s i a n a State University, Baton Rouge, L o u i s i a n a 70803 (U.S.A.) 2 D e p a r t m e n t of V e t e r i n a r y Pathology, College of V e t e r i n a r y Medicine, Texas A & M University, C o l l e g e Station, Texas 77843 (U.S.A.)

(Accepted

i0 D e c e m b e r

1980)

ABSTRACT

Snider III, T.G., Adams, L.G., and Pierce, K.R., 1981. Effect of N e o n a t a l T h y m e c t o m y and A n t i l y m p h o c y t e G l o b u l i n on N o n - S p e c i f i c Mitogen S t i m u l a t i o n in H o l s t e i n - F r i e s i a n Calves. Vet. Immunol. I mmu n o p a t h o l . , 2: 175-188.

Nine of seventeen neonatal H o l s t e i n - F r i e s i a n calves were thymectomized, treated with a n t i l y m p h o c y t e globulin, and monitored for i mmunol o g i c functional ability for 4 to 6 months. The thymus weights for 4 to 10-day-old calves and 4 to 6-month-old calves indicated a c o n t i n u e d increase in total weight. This indicated significant thymic involution had not o c c u r r e d at 4 to 6 months. Following thymectomy a wasting syndrome was not o b s e r v e d although an increased incidence of a lowly v i r u l e n t v i r u s infection did occur. A significant decrease in c ircula t i n g lymphocytes was observed. P e r i p h e r a l blood lymphocytes were stimulated in vitro by nons pecifi c mitogens, p h y t o h e m a g g l u t i n i n , bacterial l i p o p o l y s a c c h a r i d e and pokewee d m i t o g e n using the whole blood culture method. Observations included a g r e a t e r response to p h y t o h e m a g g l u t i n i n and pokeweed mitogen in summer months and v a r i a b l e age related response to all mitogens. L i p o p o l y s a c c h a r i d e s t i m u l a t i o n results were inconclusive. It was concluded that neonatal thymectomy was not a satisfactory e x p e r i m e n t a l p r o c e d u r e for the p r o d u c t i o n of selective i m m u n o s u p p r e s s i o n in the bovine species. INTRODUCTION

Neonatal serum

(ALS)

thymectomy or

the s e l e c t i v e (Miller,

1962,

(TX)

in laboratory

antilymphocyte suppression East

1973).

Neonatal

humoral

immune

TX

(HI)

globulin

of cell

et al.,

1967;

in calves response

was

mediated Iwasaki not

animals

(ALG)

and man

induce

immune

(CMI)

responses

et al.,

1967;

Lance

associated

to o v a l b u m i n

and a n t i l y m p h o c y t e

in animals

with

or tetanus

et al.,

an altered toxoid

in a

176

limited

study

et al.,

1968).

unthrifty wasting

that

observed

et

and P a p p ,

1960;

2,000

cells

in 2 o f

al.,

successful

neoplastic

cells,

(Donawick calf

to

et

TX

et

calves The

has

with

1974;

1967);

1974)

or

of

Eisen,

AND

neonatally

from

in J u n e

3-day-old a well

2 began

in the w i n t e r

scope

of

to

count

calves

necessarv

shown

the

bovine

lymphocytes

to t r e a t

of

et

et

et

was

neonatally

of CMI

responses.

mitogens

lipopolysaccharide the CMI

al.,

al.,

response

Muscoplat

and

non-sDecific

capacity

The

alloqrafts

removal

the

reduces has

derived

made

Mechaness

colostrum-fed

managed

and J u l y

(IC, g r o u p

T X was

Various

of

is r e f l e c t i v e

TX

1,000

of

to N S M

CMI

not

et

et

al.,

to the

and HI

1975).

has

Lazary

Lazary

lymphocytes

of both al.,

1973;

1973;

(LPS),

system

The been

response

of

reported.

METHODS

control

Neonatal

1974;

of

(Gabel

in c a l v e s

ALS

thymus

been

(Mackaness

respectively.

al.,

leukocyte

selectively

have

bacterial

the

1972;

(PWM),

of

has

response

system

was

Animals

Seventeen

began

the HI

with

selective

of

the

calf

29 c a l v e s

blood

skin

Studies

seeding

and

et

a decrease

or A L G

of n o r m a l

the

an

to

condition

Carroll

involving

use of ALS

treatment

(PHA)

Fitzgerald,

ALS

1975).

for

calves

1968).

The

attempt

(Carroll

similar

in a h e i f e r

although

using

a measure

(Eisen,

from

Experimental

obtained

as

mitogen

responsiveness

MATERIALS

al.,

peripheral No

1966;

in the p e r i p h e r a l

et

Bach,

or A L G

1974),

pokeweed

lymphocytes

1967),

continued

lymphocyte

accepted

(Parrott,

NSM,

al.,

bull

1967)

unthrifty

al.,

et

phytohemagglutinin

been

al.,

of

et

in 2 e x p e r i m e n t s

1973).

ALS

al.,

or

al. , 1973).

1970;

et

not o b s e r v e d

transplantation

with

al.,

degree

(NSM),

was

observed

et

but

al.,

be born

(Schultz

(Coleman

(Carroll

(Lance

of CMI r e s p o n s e s

and a t r a n s i e n t

immunosuppression

function

allowed

calves

ul was

3 TX s a l v e s

(Leighton

condition

1967)

evaluation

of 4 H o l s t e i n - F r i e s i a n

rodents

Chanana

per

Biological CMI

study

in T X

wasting

(Leighton

include

developed

in 7 TX bull This

not

In the

condition

syndrome

1968).

did

4)

with

at 4 to

experimental

this

parenthesis):

paper.

and

4 TX

10 days

lasted (group and

procedures

These

prescapular

Holstein-Friesian in 2 c h r o n o l o g i c a l

5 neonatally

calves with

dairy

lymph

two

node

(qroum

I) and

were

included

TX

2)

approximately 4 IC

calves

the

biopsy

and

(group

and

ages

of

(5 w e e k s

calves

3)

Phase

calves. s h a m TX.

beyond

calves and

I

4 intact

4 were

are

were

Phase

5 months.

in g r o u p

performed

(with

bull

phases.

the

given

in

approximately

177

12 weeks), diarrhea

chicken

virus

vaccination

and

respectively) The

results

paper will

of

be identified

calves with 3 were

were

equine

and

and

and severe

erythrocyte activity

(11 and

be reported

antibovine

was

with

viral abortus

15 weeks, (16-24 weeks).

separately

and

in this

procedures.

titers

with was

produced

of

the g l o b u l i n

4 times

ALG

horse

assayed

in vitro 1:128

bovine

transient

lymphocytes

in an 8-month-old

1:64,

respectively,

for

The ALG a 50% d e c r e a s e

in hematocrit

at 72 hours.

erythrocytes.

at 24

The ALG

The

was

anti-

of the a n t i t h r o m b o c y t e

thrombocytopenia

was observed

anti-

lymphoagglutination

calf with

independently

2 calves

The

(2 gm/dl).

50% decrease

washed

removed

and

(NHG).

for

and

fraction

(4,000/ul)

at 48 hour

(0.04 g/kg)

globulin

in an 8-month-old

thrombocytopenia

blood

injected

normal

in vivo

absorbed

which

(0.04 g/kg)

bovine

Brucella

inoculation

immunological

approximately

activity

of p e r i p h e r a l

with

for

assayed

subsequently

will

as other

activity

1:16

in lymphocytes, hours

immunization

produced

lymphocytotoxicity

(0.025 g/kg)

2 only),

Calmette-Guerin

subcutaneously

treated

lymphocyte

the p l a s m a

(7 weeks),

in phase

Globulin

intervals

and

toxoid

these p r o c e d u r e s

in grou p bovine

tetanus

immunization

(9 weeks

and B a c i l l u s

Antilymphocyte

All TX

erythrocyte

infection

and

8 hours

a 73% reduction

after

I dose

calf.

Thymectomy

The TX p r o c e d u r e the xiphoid thymus

and all

building.

From

were

commercial building Routine

milk

on

calf

1967).

surgery

was

Mean

from other

replacer

concrete

floors

covered

The

in stalls with

fresh

of measuring

were

and analyzed

total

leukocyte

obtained

from

for d i f f e r e n c e s

(TWBC)

related

until

were

necropsy

were

pine wood

and daily

.05 g)

lymphocyte collections

to TX.

frame

shavings.

temperature, u and

the

fed

in an enclosed

(400,000 and

procedures

calves

rectal

I to 4 blood

to

of the thoracic

the e x p e r i m e n t a l

at age 3 days

of p e n i c i l l i n - s t r e p t o m y c i n

weekly

from the mandible

surgical

within

animals.

and housed

consisted

incision

for removal

Standard

performed

the day of arrival

monitoring

concentrations

a midline

thoracotomy

et al.,

isolated

administration weeks.

involved

a sternal

(Leighton

observed

calves

with

for 4

(TLYM) from each

178

Lymphocyte

Upon

Blastogenesis

the arrival

blood was

of each

collected

(5-10 u/ml of blood) using al.,

the whole 1974).

for

Oxnard,

blood

culture

mixed ing

with

USA).

penicillin

no mitogen

Laboratories,

E. coli,

in phase

2, Difco

Company,

Grand

manufacturer The

were

50 ul of RPMI-1640 Thymidine Nuclear,

cultures were

were

sample The

filters

were

Ill.,

stimulation triplicate cultures The

USA).

culture

Statistical

IDepartment

added

with

Counter,

counts

were

stored

a liquid Packard counts

per minute

in the unstimulated using

System's The

study

USA.

analysis

well,

before

New

the

the cultures multiple

USA).

and placed

in

(29 g Fluoralloy

in the dark

and

per

at 4C until spectrometer

Company,

Downers

lymphocyte the

in the NSM

control

designed

Statistics,

(3H-TdR,

fluid

average

of

stimulated

cultures.

of v a r i a n c e

(SAS) general was

thymidine

Wis.,

made

for After

by dividing

(CPM)

air

collection.

scintillation

were

by the

= 6.7 Ci/mmol,

Instrument

calculated

055:B5,

Biological

a semi-automatic

for 24 hours

Difco

LPS

E. coli,

to each

16 hours

10 ml s c i n t i l l a t i o n

of E x p e r i m e n t a l La.,

activity

Madison,

(SI) were

Analysis

Baton Rouge,

specific

at 70C

(40 ug/ml,

of 5 ul/ml.

USA) was

Hiller,

contain-

as recommended

blood

Otto

The v i a l s

analyzed

IBM 360 computer I.

after

was

either

in 5% CO 2 h u m i d i f i e d

hour

an additional

Five minute

by the CPM

SI were

concentration

and was

of wells

which

Island

et

to

Plastics,

1974),

diluted

using

was m e a s u r e d

indices

(Grand

was

filters

dried

of toluence).

et al.,

fiber

with

(TriCarb S c i n t i l l a t i o n Grove,

for

PWM

(RPMI-1640

PHA-P

0.5 uCi of tritiated

Mass.,

(SAMSH,

vials

radioactivity

which

(methyl-3HO), Boston,

media

Muscoplat

of blood

coli,

I or 50 mg/ml,

at 37C

containing

1972;

(Falcon

following:

(Lazarry

one

blastogenesis

sample

streptomycin)

or PWM

USA)

within

plates

culture

in phase

incubated

on glass

harvester

scintillation gallon

NY,

incubated

collected

tissue

USA)

and Good,

in each

thereafter,

heparinate

in each of a series

100 ug/ml

at a final

beginning

England

of

Mi.,

of lymphocyte

from E s c h e r i c h i a

microtiter

and

intervals

into sodium

(Park

ul of blood

Laboratories),

and used

NET-027

LPS

0111:B4

Island,

cultures

30-32 hours

method

or one of the

Detroit,

(0.05 ug/ml,

at 2 weeks

lymphocytes

using

Fifty

an equal v o l u m e

100 u/ml

contained

of

(PHA-P),

in triplicate Ga.,

and

the m e a s u r e m e n t

The response

phytohemagglutinin-P measured

calf

by jugular v e n i p u n c t u r e

linear

(ANOVA) models

with

the

program

on an

as a 2 X 2 factoral

Louisiana

State University,

179

arrangement and

no TX

mental and

of

treatments

and p h a s e

hypotheses

IC c a l v e s

2) t h e r e

was

in p h a s e

to the

responses blood

the by

same

were:

I (group

to NSM,

used

lymphocytes the

tested

no d i f f e r e n c e

of

the

in a s p l i t

I and p h a s e

for

per

procedure

to

2 and

and p h a s e

4)

I and

assays

into

test

on

was

the p e r i p h e r a l

(LY)

design.

a split

I) t h e r e

in p h a s e

the N S M

ul

plot

2 with

phase

an

the P H A - P

the

factors The

no d i f f e r e n c e

blood

and

The time.

2 (groups

lymphocyte

CPM

between

2 calves.

index

TX

I and

3)

In a d d i t i o n counts

created

(PHA/LY)

were TX

experi-

per

ul

by d i v i d i n g

were

anlayzed

hypotheses.

RESULTS

The the

weight

narrow

ooint

of

thymic from

of

the

isthmus

division

tissue

from TX

respiratory

ratio

(CTR) v a r i e d

TABLE

from

IA IB IC ]D 2A 2B 2C 2D 2E OA OB Mean n

of

0.14 the

thymic

tissue and

calves

to

139.3 g from

to

f r o m each at the

thoracic

including

immediately (Table

1.49

to 2.86

.035.

Similar

IC c a l v e s

revealed

and

inlet The

used

mean

(OA and OB)

surgery cervial

was to

and

thymic

with as the

weight which

of

a

thymus

body

weight

calculations

weight

of

died

97.8 g w i t h

thoracic

the p e r c e n t a g e

measurements a mean

measured

thymus.

2 calves

The

was

thoracic

after

I).

calf

after

of 2 7 2 . 3 g w i t h

I

T h e ages Friesian

Calf

37.6 g

removed

cervical

acidosis

of

necroosy

of of

ranme

varied

thymus

at the calves

Age (days) 8 7 4 7 7 10 9 8 7 6 4 7 11

time

of

thymectomv

and

thymic

T h y m i c W e i g h t (g) Cervical Thoracic Total 22.5 81.0 90.0

15.1 37.0 31.5 .... 57.6 23.8 71.4 32.5 64.7 39.5 61.6 23.6 98.0 41.3 73.0 30.5 . . . . . . . . 68.9 30,5 9 9

* R a t i o b e t w e e n the w e i q h t n = n u m b e r of calves.

of

cervical

37.6 118.0 121.5 84.3 81.4 103.9 104.2 85.2 139.3 103.5 97.0 97.8 11

and

size

CTR*

for

11 H o l s t e i n -

% Body

1.49 2.19 2.86

0.14 0.30 0.31 0.21 0.18 0.23 0.27 0.18 0.35 0.27 0.25

2.42 2.20 1.64 2.61 2.37 2.37 .... 2.26 9

thoracic

Weight

thymus.

180

a mean CTR of ( T a b l e II).

TABLE

2.17

and

a slightly

Calf

at the calves

time

Age (days)

3A 3B 3C 3D 4A 4B 4C 4D Mean n

of

euthanasia

23 22 21 19 19 18 17 17 19.5 8

Tissue

258.2 168.4 212.8 214.1 176.3 117.9 181.2 162.5 186.4 8

suspected

cervical

and

Residual

thymic

2B and

113.6 91.7 85.8 82.3 71.6 69.5 84.8 87.5 85.9 8

of

to be

thoracic

thymus

regions

tissue,

was

of

identified

located

in the

larynx

and

closely

associated

glands.

in an IC calf

Thymic

(calf

in all

All T X

the

body

weight

s h a m TX

calves

and w e r e

at rest

to s u r g i c a l

infected

coli)

and

treatment

calves) mucosa

with

focal

systemic

virus

made

thymus.

the

each

TX

anterior calf.

in c a l v e s

region,

submandibular

located

dorsal

structure

uneventful mild

infections

of

most

of

erosions

calves

and

and h y p e r e m i a persisted

disease

isolation infection

for

being were

in 5 of

IA,

ID,

2A,

dorsolateral and/or to the the

to

Darotid larynx

thymus

and

was

shallow of

the

as long

anorexia

consistent

(Griesemer

despite

and C o l e ,

"7 o f of

8 IC

the o r a l

fungiform

4 weeks

in calf with

topical

peroxide.

9 T X and

lingual

These

Klebsiella,

ulcerations

as

surgery

3B.

bovine 1960).

with

in

recumbency

9 TX c a l v e s .

hydrogen

(6 of

from

Sternal

(Proteus,

in 3 c a l v e s

(Lugol's) the

recoveries

replacer.

bacteria

persisted

solution

lesions

and v i r a l

stomatitis

also

to n u r s e

wound

infection

hypertrophy The

of

was

cervical

Weight

0.31 0.20 0.20 0.26 0.17 0.16 0.21 0.23

from of

histologically

the

% Body

2.27 1.84 2.84 2.60 2.46 1.70 2.14 1.86 2.17 8

thoracic

histological

opportunistic

iodine

developed

evidence

able

2 to 3 w e e k s

papillae.

lesions

with

the

with

Within

The

CTR*

necropsy

with

for 8 H o l s t e i n -

calves.

and

lead

and

anterior

tissue

3D).

I to 5 h o u r s

became

size

collected

during

was

normal

thymic

371.8 260.1 298.6 296.4 247.9 187.4 266.0 250.0 272.3 8

cervical

2C,

salivary

and

T h y m i c W e i g h t (g) Cervic@l Thoracic Total

* R a t i o b e t w e e n the w e i g h t n = n u m b e r of c a l v e s .

E.

percentage

II

T h e ages Friesian

the

lower

the o n l y

Histological papular

181

The and

average

associated TLYM

selected was

for T X

and

(Figure

TWBC

the TX TX

The

the TX when

and

in the T X

and

from

in the

were

The

calves

was

to 4054

mean TLYM

For w e e k s

was

and

calves

in the T X

(Figure

I).

were

not g r e a t l y

distinctly

TLYM

of

the

4-17

2840)

calves

between

ages

I

different

2 groups

of

than

mean

ages.

until

lower

TLYM

in

in the

The TLYM

the

third

occurred

in

in the TX

The g r e a t e s t

calves

the m e a n

the m e a n T W B C

(P < 0.05)

at all

lower TLYM.

the 4th week, between

more

The w e e k l y

IC c a l v e s

had

until

significantly

in the IC c a l v e s

mean

age

mean TWBC

between

difference

(mean o f

than

in the w e e k l y

(TWBC)

for

calves

mean TWBC

higher

(P < 0.001)

IC c a l v e s .

1242

IC c a l v e s

calves

The

the

in the IC c a l v e s

lower

the T X

of

t-test

overall than

leukocytes by A N O V A

of TX.

arrival

Student's

lower

significant

overall

than

calves

using

of

blood

analyzed

I).

in the T X

IC c a l v e s v a r i e d

total and

the e f f e c t

time

IC g r o u p s .

the T W B C

a highly

to

the

(P < 0.05)

17 w e e k s

calves.

related at

of

determined

no d i f f e r e n c e s

Although

weekly

were

collected

significantly

there was

concentrations

(TLYM)

differences

in b l o o d

demonstrated

to

weekly

lymphocytes

of

week

difference at

age

8

weeks. The by

approximate

the o v e r a l l

7491CPM CPM.

mean

for PWM.

Individual

marked

to N S M

differences III,

(Tables

for P H A - P ,

mean CPM

for

seldom varied the v a r i a n t

related

the o t h e r

ages.

the w e e k l y

mean

LPS

the m e a n s

was

SI of

were

and

by N S M

was

reflected

1846 CPM

control

for LPS

cultures

by more

than

culture

result

weekly

mean

0.003)

different

PHA-P

there

was

which

implies

the

not

was

and

911

20% but w h e n was

not

used

to

SI

that

response

week

did

not

to PHA-P,

(P < 0.003)

LPS o r

in m e a n s

I SI was g r e a t e r

(P < 0.02)

age g r o u p s

mean NSM

VI,

VII, was

than

interaction

at

between

which

implied

the d i f f e r e n c e s

SI due

to calf

age w a s

and

(phase

lower

the

I plus

(P < 0.01)

the m e a n

PHA-P

between

VIII).

at the d i f f e r e n t

a significant

in the

a difference

the m e a n

different

higher

I to 7 w e e k s

consistent.

the m e a n L P S

and PWM w e r e

was

ages

significant

in the w e e k l y

(Tables

for

to T X

There

(P < 0.05)

calves

higher

related

to age w h e r e There

The variation significantly

stimulation

IV, V).

for PHA-P

PHA-P

34323 CPM

of

did o c c u r

PWM

(Winter)

induced

overall

cultures

demonstrate

were

stimulation

the SI.

response

between

of

counts The

deviations

calculate The

degree

mean

The

I (summer)

mean

in p h a s e LPS was

phase ages

phase

SI

I. lower

2) w e r e

interaction

for P H A - P The

weekly

in p h a s e

and,

between

2

and P W M mean The

(P <

in a d d i t i o n ,

phase

I and p h a s e

phase

I.

significantly

(Table VIII)

SI of p h a s e

and

and

age,

2 calves

were

182

X 103 13

12, I I0

9 8

V ',d

7'

fs

"'"- "--,. / •

;I

6'

;

5

%

",,,

S"

\

,/% ~e

"%,.,%

,# % p l "%"e"o ~ ' ~

4

I c -WBC

..... IC - LYM ....

T X - WBC

TX - LYM . . . . . .

WEEKS

Fig. I. Mean thymectomized

TABLE

w e e k l y l e u k o c y t e (WBC) and l y m p h o c y t e (TX) and intact c o n t r o l (IC) calves.

(LYM)

counts

in

III

The m e a n p h y t o h e m a g g l u t i n i n - P stimulation indices for l y m p h o c y t e s in w h o l e blood f r o m t h y m e c t o m i z e d (TX) and intact c o n t r o l (IC) calves in p h a s e I (PI) and p h a s e 2 (P2). Treatment Age

G r o u p ns

(weeks)

I 2,3 4,5 6,7 M e a n ns n

I(TX,P2)

2(TX,PI)

3(IC,P2)

4(IC,PI)

Mean*

35.98 57.15 23.88 30.88 36.19 4

158.32 70.05 33.76 28.99 72.23 5

36.03 21.86 15.98 32.80 24.96 4

130.02 56.92 43.22 52.27 69.09 4

94.10 53.03 27.33 35.54

* D i f f e r e n c e in m e a n s b e t w e e n ages, s i g n i f i c a n t , P < 0.0003. n s D i f f e r e n c e b e t w e e n means, not s i g n i f i c a n t , P > 0.05. n = n u m b e r of calves.

17

183

TABLE

IV

The mean in w hol e in phase

E. coli l i p o p o l y s a c c h a r i d e stimulatin indices for lymphocytes blood from t h y m e c t o m i z e d (TX) and intact control (IC) calves I (PI) and phase 2 (P2). Treatment

Age

Group*

(weeks) I(TX,P2)

2(TX,PI)

3(IC,P2)

4(IC,PI)

2.06 2.69 1.82 3.38 2.42 4

1.20 0.75 0.83 1.20 0.95 5

4.36 1.49 1.86 2.47 2.43 4

1.32 1.43 1.23 1.03 1.27 4

I 2,3 4,5 6,7 Mean ns n

2.17 1.42 1.48 2.03 17

* D i f f e r e n c e in means related to age, thymectomy and phase significant, P < 0.02. n s D i f f e r e n c e between means, not significant, P > 0.05. n = number of calves.

T A BLE

Mean ns

interaction,

V

The mean p o k e w e e d m i t o g e n s t i m u l a t i o n indices for lymphocytes in whole blood from t h y m e c t o m i z e d (TX) and intact control (IC) calves in phase I (PI) and Phase 2 (P2). Treatment Age

G r o u p ns

(weeks) I(TX,P2)

2(TX,PI)

3(IC,P2)

4(IC,PI)

5.74 6.09 4.80 6.73 5.70 4

9.68 21.44 10.18 6.92 14.30 5

12.44 4.09 3.93 5.09 6.20 4

30.44 15.14 14.50 8.20 16.85 4

I 2,3 4,5 6,7 Mean ns n

n s D i f f e r e n c e between means, n = number of calves. not

consistently

did not differ phases. calves

When

different

the weekly

between

ages

significant

(P < 0.0001)

The each

(Table

with

ages.

the d i f f e r e n c e

The weekly

the P H A / L Y

were

tested

17

mean LPS or PWM in either

indices

SI

of the 2

of TX and IC

for s i g n i f i c a n t

in mean LY between

difference

14.90 13.34 7.76 6.84

P > 0.05.

age of the calves

mean LY and

I to 7 weeks

only

significant,

at all

significantly

differences

phases

not

Mean ns

in mean PHA/LY

ages were indices

between

IX).

mean SI for PHA-P,

phase were

tested

LPS

and PWM

for s i g n i f i c a n t

for each

treatment

differences

over

group the

within

first

21

184 TABLE

VI

The mean p h y t o h e m a g g l u t i n i n - P w h o l e blood from p h a s e I (PI)

Age

(weeks)

stimulation indices for l y m p h o c y t e s and p h a s e 2 (P2) calves.

PI

P2

PI-P2*

in

Mean +

I 145.74 36.01 109.73 94.10 2,3 65.12 38.15 26.97 53.03 4,5 37.96 20.49 17.47 27.33 6,7 39.34 31.73 7.61 35.54 Mean/ 70.92 30.83 n 9 8 17 * D i f f e r e n c e b e t w e e n m e a n s at v a r i o u s ages, s i g n i f i c a n t , P < 0.01. +Difference in m e a n s b e t w e e n ages, s i g n i f i c a n t , P < 0.003. ~Difference in m e a n s b e t w e e n e x p e r i m e n t a l phases, s i g n i f i c a n t , P < 0.05. n = n u m b e r of calves.

TABLE

VII

The m e a n in w h o l e

Age

E. coli l i p o p o l y s a c c h a r i d e s t i m u l a t i o n indices for b l o o d from p h a s e I (Pl) and p h a s e 2 (P2) calves.

(weeks)

I 2,3 4,5 6,7 Mean* n * D i f f e r e n c e in m e a n s P < 0.05. nsDifference between n = n u m b e r of calves.

TABLE

PI 1.25 1.00 1.01 1.13 1.08 9 between means,

P2 3.21 2.03 1.84 3.04 2.42 8 experimental not

P I - P 2 ns

M e a n ns

-1.96 -1.03 -0.83 -1.91

phases,

significant,

2.17 1.42 1.48 2.03

17 significant,

P > 0.05.

VIII

The mean p o k e w e e d m i t o g e n s t i m u l a t i o n indices for blood from p h a s e I (Pl) and p h a s e 2 (P2) calves.

Age

lymphocytes

(weeks)

I 2,3 4,5 6,7 Mean* n * D i f f e r e n c e in m e a n s nsDifference between n = n u m b e r of calves.

PI

P2

lymphocytes

P I - P 2 ns

in whole

Mean ns

21.54 9.09 12.45 14.90 19.07 5.00 14.07 13.34 12.10 4.51 7.59 7.76 7.49 6.11 1.38 6.84 15.42 5.92 9 8 17 b e t w e e n e x p e r i m e n t a l phases, s i g n i f i c a n t , P < 0 . 0 5 means, not s i g n i f i c a n t , P > 0.05.

185 TABLE

IX

The

mean P H A / L Y *

Age

(weeks)

indices

in phase

Phase

I (PI)

and phase

I

Phase

2 (P2)

calves.

2

Mean ns

I 10.56 2.41 6.73 2,3 8.39 3.52 6.20 4,5 8.45 4.04 5.77 6,7 7.80 4.70 6.25 Mean + 8.73 3.72 n 9 8 17 *PHA/LY = counts per minute of p h y t o h e m a g g l u t i n i n - P stimulated lymphocytes in whole blood divided by the total blood lymphocyte concentration. + D i f f e r e n c e in means between experimental phases, significant, P < 0.0001. n s D i f f e r e n c e between means, not significant, P > 0.05. n = number of calves.

weeks

related

response I the

difference

signifi c a n t which

only

to TX.

to PHA-P,

in mean PHA-P

although

implies

that

ages,

similar

age related

SI

no effect

although

various

LPS

In phases

LPS or PWM was

(P < 0.0001)

related

the response did

differences

and PWM

related

(P > 0.05)

TX and IC calves

(P < 0.05)

I and 2 no difference

demonstrated

not respond

between

(P < 0.0004)

means

In phase

to age was

of treatment to mitogens

in the

to TX.

was

were

demonstrated different

differently. for PHA-P

at

In p h a s e

2

(P < 0.0002),

SI were observed.

DISCUSSION

Neonatal

TX was

the thymus. surgical

procedure

evidenced

of

surgical calves

of

the

and

that

N eon a t a l alterations

TX

and

thymus

and TLYM was

variation

cervical appears

prior

have

did not

attributed

consistent in calves,

with mice

thymic

cause

the effect

of

As

the close complete

by c o n v e n t i o n a l

weight

of neonatal indicated

to grow

the severe

with

of

that

up to this

age

involution.

neonatal

to TX did occur.

rats

and

(272.3 g)

continues

associated

and

thymus.

glands,

thymic

calves

to normal

been

of the

to be impossible

calves

portion

to be a d i f f i c u l t

salivary

in mean

17 to 23 w e e k - o l d

was

of the major

found

in 5 of 9 TX calves

difference

in the calf

which

changes

TX

thymus

of H o l s t e i n - F r i e s i a n

necropsy

the removal

calf was

to anatomic

The

and

for

in the

larynx

the bovine

(97.8 g)

thymus

neonatal

due

methods.

the

although

TX

by the residual

association removal

successful

Neonatal

The

immunological TX in rodents

depression

in utero TX of

(Miller,

1962;

Miller

in TWBC

lambs

and

and Osoba,

186

1967;

Carroll

evidence

of

(Leighton sternal the

et

al.,

et

al.,

surgical

normal

localized

1968;

a wasting

1967)

or

wound

sternal areas

the

skin

No variations

infections,

nor

differently

than

differences

case

phase, 1978)

TX

explanation

was

found

of

the

and

at

as

with

residual

TX

rest

The

aggravated

were

were

no

calves

TX.

and

low g r a d e

5 calves

confined

by to

suppurative

attributed

thymus

to

these

respond

calves.

to N S M

and

IC

did

not

calves.

than

lower

In

phases

a different

the

calves

was

in

after

incited

results

other

for

1962)

were

subcutis

experimental

where

(Miller, which

There

described

test

other

between

1971).

in N S M

responses

of LPS,

no

the

the

between

a difference the

did

lymphocyte

rodents

recumbency

of

and M o r r i s ,

as p r e v i o u s l y

infections

processes.

The

Cole

syndrome

LPS

demonstrate almost

all

unrelated

seasonal

analyses

to TX.

preparation

responses

was

influences

during

consistent

the

there

Except

used

in each

(Soper

winter

was

in

et

al.,

months

(Phase

2). The

principal

the v e r y for

the

PHA/LY were

high

age

level

related during

differences indices,

in

no

indices

between

The

phase

I and

PHA/LY

significant

the

mean

the

2 calves

PHA-P

PHA-P

concentration

phase

of

the

when

in m e a n

highly

responsiveness

in

I and,

lymphocyte

differences

demonstrated.

a greater

variation

week

by

SI

CPM

the

was

were

due

to

adjusted

creation

of

indices

related

to TX

difference

in mean

PHA/LY

uo

lymphocytes

to

age

7 weeks

in p h a s e

indicated

2 calves

to N S M

21

in

stimulation. The vitro phase both

difference

biweek!v

of

2 calves

to P H A - P ,

in

The

the

did

Marked

higher

the

greater

in

LPS

and

the

induced

by

neonatal

whole

significantly

obtained

with

response

of

summer

PHA-P

months

reflected

TX

and

and

I to

the

PWM

as m e a s u r e d

degree method.

a brief

regimen

lymphocyte

response

blood

weeks

to P H A - P

culture

in v i t r o

in N S M

peripheral

ages

I calves

blood

alter

differences

for

phase

PWM

in

not

the

from

and

seasonal

values

addition

intevals

lymphocytes

calves

changes

treatment NSM.

at

response

in

were

summer

*H-TdR

the

LPS

in

and

of v a r i a t i o n

of

ALG

responses

observed

months.

lymphocytes bv

and

to

with

In

to P H A - P

was

much

uptake.

ACKNOWLEDGEMENTS

The

authors

technical This the

work

Texas

wish

to

assistance was

thank

and

supported

Agricultural

Betty

Connie

by N I H

Experiment

Rosenbaum

Leggett

for

Fellowship Station.

and R o n manuscript Grant

Blakely

for

preparation.

5T01-GM01232-13

and

187

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