Background: The colostrum of HIV-infected mothers contains a high number of HIV copies and is considered highly infectious.

Furthermore it contains large numbers of macrophage and other mononuclear cells that are known to incorporate virus. While

prevention protocols in Western countries suggest the interruption of breast feeding, at least for the first few months of life, this

practice is not advisable in developing countries.

Methodology: The aim of this study was to determine the HIV load and the concentrations of IL-18, IL-16, IL-12, TGF-beta1 and

TGF-beta2 in the colostrum of HIV-infected mothers living in Burkina Faso. The women all received nevirapine prophylaxis

during labour.

Results: The viral load in the colostrum decreased rapidly during the first three days following delivery, while the concentration of

IL-18 and IL-16 increased in the same period. IL-12, TGF-beta1 and TGF-beta2 did not show significant variations in the first

three days after delivery.

Conclusions: Since the viral load decreases in the colostrum of nevirapine-treated expectant mothers, our data suggest single

dose nevirapine combined with interruption of early feeding may have potential as a way to reduce the risk of MTCT

Castronuovo, Piero

Musumeci, Maria

Musumeci, Salvatore

Palano, Maria

Pietra, Virginio

Simporé, Jacques

UnissResearch

Original Article  
Human Immunodeficiency Virus Type-1 and cytokines in colostrum from 
HIV-infected mothers in Burkina Faso   
Maria Musumeci1, Grazia Maria Palano2, Piero Castronuovo3, Virginio Pietra4, Jacques 
Simpore4, and Salvatore Musumeci5.  
1Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy;   
2Department of Paediatric, University of Catania, Italy;  
3Clinical Pathology Service, M Ascoli-Tomaselli Hospital, Catania, Italy;  
4Centre Medical Saint Camille (CMSC), Ouagadougou, Burkina Faso, University of Ouagadougou, UFR/SVT, Burkina Faso;  
5Department of Pharmacology, Gynecology and Obstetric, Pediatrics, University of Sassari, Italy, Institute of Biomolecular 
Chemistry, CNR, Li Punti (SS), Italy.   
Abstract 
Background: The colostrum of HIV-infected mothers contains a high number of HIV copies and is considered highly infectious. 
Furthermore it contains large numbers of macrophage and other mononuclear cells that are known to incorporate virus. While 
prevention protocols in Western countries suggest the interruption of breast feeding, at least for the first few months of life, this 
practice is not advisable in developing countries.  
Methodology: The aim of this study was to determine the HIV load and the concentrations of IL-18, IL-16, IL-12, TGF-beta1 and 
TGF-beta2 in the colostrum of HIV-infected mothers living in Burkina Faso. The women all received nevirapine prophylaxis 
during labour.  
Results: The viral load in the colostrum decreased rapidly during the first three days following delivery, while the concentration of 
IL-18 and IL-16 increased in the same period. IL-12, TGF-beta1 and TGF-beta2 did not show significant variations in the first 
three days after delivery.  
Conclusions: Since the viral load decreases in the colostrum of nevirapine-treated expectant mothers, our data suggest single 
dose nevirapine combined with interruption of early feeding may have potential as a way to reduce the risk of MTCT. 
Key Words: HIV, colostrum, Burkina Faso.   
J Infect Developing Countries 2007; 1(1):25-29.   
Received 24 January 2007 - Accepted 1 June 2007.  
Copyright © 2007 Maria Musumeci et al. This is an open access article distributed under the Creative Commons Attribution License, which 
permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.   
Introduction 
Human colostrum is a complex liquid produced 
by the mammary glands to sustain the newborn in 
the first days after delivery. After the first few days 
the composition of the mammary gland secretion 
changes to become a source of nutriment, but 
during the first 3 days after delivery the principal 
role of colostrum is to provide immunity to the 
newborn [1,2] as it contains numerous 
lymphocytes and macrophages. Changes that 
occur during this transformation include decreases 
in the concentration of some components, 
including IgE, prolactin and endorphin [3,4] and 
increases in the concentration of other 
components such as fucosyl and other 
oligosaccarides [5]. The concentrations of the 
macrophage enzyme chitotriosidase [6], as well as 
some cytokines (IL-1beta, IL-2, IL-6, IL-8, TNF-
alpha) decrease rapidly during the first three days, 
suggesting that mononuclear cell numbers 
decrease, in conjunction with increasing 
concentrations of IgA and IgG [7-9] . 
In mothers infected with human 
immunodeficiency virus (HIV), the expression of 
some cytokines (such as TGF-beta) in colostrum 
may promote HIV replication in mammary 
epithelial cells [10] leading to high colostral viral 
load and increased risk of mother to child 
transmission (MTCT) through breastfeeding. While 
in some countries HIV-infected mothers are 
advised not to breastfeed their infants in order to 
reduce this risk [11,12], however, such advice is 
regarded as unsuitable for mothers in developing 
countries because of the increased risk of infant 
Musumeci et al - HIV load in colostrum    
26 
J Infect Developing Countries 2007; 1(1): 25-29. 
diarrhoea through unhygienic preparation of milk 
formula.  
The use of single dose nevirapine, and other 
anti-retroviral therapies, during pregnancy has 
been shown to significantly reduce the risk of 
MTCT of HIV [13,14]. Preliminary studies have 
shown that the viral load of colostrum from HIV-
positive mothers, treated with a single dose of 
nevirapine, fell within the first three days following 
delivery from an initial load of >100,000 copies/ml 
to <10,000 copies/ml [11]. Colostrum may 
therefore be a significant risk for MTCT of HIV and 
a temporary suspension of breastfeeding, for three 
days after birth, may significantly reduce the risk of 
transmission. The risk of contamination of milk is 
low, as milk can be safely prepared if delivery is 
carried out in a health-care setting. Following this 
temporary suspension, breastfeeding may be 
continued and the risk of infant diarrhoea is limited.  
In this study we confirm a rapid reduction of 
viral load in colostrum from mothers treated with 
single dose nevirapine, and also report on the 
variations in cytokine concentrations that may 
affect viral load. These studies form the basis for 
potential interventions that may determine the 
efficacy of interrupted breastfeeding in prevention 
of MTCT of HIV.  
Materials and Methods 
Twenty HIV-infected mothers were recruited 
from the Voluntary Counseling and Testing (VCT) 
program at Centre Medical St Camille (CMSC), 
Ouagadougou, Burkina Faso, which provided 
ethical review and approval. Mothers with 
detectable viral loads were informed of the 
objectives of the study and following informed 
consent were allocated to receive Nevirapine (200 
mg, single dose) during labour. Nevirapine 
(2mg/Kg) was administered to their babies within 
72 hrs, following the recommended practice of the 
MTCT prevention programme at CMSC.  
All of the mothers consented to the interruption 
of breastfeeding for the first three days after 
delivery, while the nurses prepared and 
administered a 5% glucose solution for the 
hydration of the newborns. Six hours after delivery, 
colostrum was collected by delicate vacuum 
pumping, using graduated sterile tubes. The 
colostrum samples were taken to the CMSC 
laboratory where the tubes were stored at -80°C.  
Infants were fed formula (10 g each meal for 
the first day seven times a day, and increasing by 
10 g each meal for the remaining two days). 
Colostrum was collected daily from both breasts 
and discarded after representative samples had 
been stored as above. On the fourth day the 
mothers were encouraged to breastfeed their 
babies normally.   
Determination of viral load 
The HIV load was measured in serial 
colostrum samples, transported on dry ice to the 
Laboratory of Virology, Ascoli-Tomaselli Hospital, 
Catania, Italy. After thawing, colostrum samples 
were centrifuged at 680 X g for 10 min at 4°C. The 
supernatant was removed and re-centrifuged at 
10,000 X g for 30 min at 4 C° to allow removal of 
the lipid layer and cellular elements. Aliquots 
(250ul) were stored in Eppendorf tubes at -20°C. 
The HIV load was determined using the 
AMPLICOR Reverse Transcriptase Polymerase 
Chain Reaction (RT-PCR) assay (Roche). The 
results are expressed as HIV RNA copies/ml on a 
log scale.   
Determination of cytokines contents 
Cytokine concentrations (IL-18, IL-16, IL-12, 
and TGF-beta 1 and 2) were determined in the 
same serial colostrum samples, with a commercial 
enzyme-linked immunosorbent assay (R&D 
Systems, Milan, Italy).    
Statistical analysis 
The non-parametric Mann-Whitney U test was 
used for statistical analysis.  
Results 
The HIV load in the colostrum of 20 HIV-
infected mothers (aged 15-44 years) ranged from 
8,000 to 120,000 copies/ml (mean 80,200 ± 
72,600 copies/ml). By the second day the HIV load 
had decreased significantly (see figure 1) to a 
maximum of 35,000 copies/ml (P<0.0001), and on 
the third day the HIV load decreased again to 
below 2,000 copies/ml (P<0.0001). 
By way of contrast, the colostral concentration 
of IL-18 and IL-16 increased over the first three 
days following delivery, though the variation was 
quite high.  
The statistical analysis of cytokine 
concentration in the first three days after delivery 
Musumeci et al - HIV load in colostrum    
27 
J Infect Developing Countries 2007; 1(1): 25-29. 
confirmed that only the concentrations of IL-18 and 
IL-16 increased significantly in the second and 
third days (Table 1), while the concentrations of IL-
12, TGF-beta1 and TGF-beta2 remained stable 
over the whole period.    
Table 1. Concentration (pg/ml) of IL-18, IL-16, IL-12, 
TGF-beta1 and TGF-beta2 in the colostrum from 20 
HIV-infected mothers.   
Cytokines 
[pg/ml, median 
(range)] 
1° day 2° day 3° day 
IL-18  
(n=20) 
10.67 
(6-18)* 
80.22  
(11-257) 
72.85  
(10-197) 
IL-16  
(n=20) 
74.35  
(25-98)* 
134.00 
(37-236) 
226.05 
(58-487) 
IL-12  
(n=20) 
38.35  
(28-62) 
31.35  
(24-34) 
30.65  
(22-34) 
TGF-beta1 
(n=20) 
66.97  
(59-71) 
82.37  
(66-106) 
69.42  
(50-78) 
TGF-beta2 
(n=20) 
912.05 
(90-2602) 
789.47 
(557-835) 
753.05 
(436-1026) 
*1° day  2° day, P<0.05.  
  
Figure 1. HIV load (copies/ml) in colostrum from 20 
HIV-infected mothers.  
Discussion 
The results clearly show that the HIV load in 
the colostrum of nevirapine-treated mothers 
decreased significantly over the first three days 
following delivery, reducing the risk of MTCT in 
breast milk. This reduction in HIV load in colostrum 
could be explained as resulting from reduced 
replication of HIV in the presence of nevirapine. 
While cell counts were not done, the decrease in 
concentration of macrophage-derived enzymes [6] 
is suggestive of a progressive reduction in 
numbers of macrophages. Macrophages and other 
mononuclear cells may be an important source of 
HIV infection, independent of the infection derived 
from free HIV virions as detected by RT-PCR [15-
17]. Other studies have suggested that cell-free 
HIV-1, which is present in mature breast milk but 
only at low concentrations [18] may not represent 
a significant source of infection in the absence of 
inflammation. Following treatment with nevirapine, 
concentrations of HIV-1 may be even lower, since 
nevirapine is excreted in breast milk.  
Factors that influence inflammatory responses 
in breast milk, such as mastitis, may also promote 
HIV transmission through the production of HIV-
infected mononuclear cells that are ingested with 
the milk [19-21]. Pro-inflammatory cytokines may 
be used as a surrogate marker of infectivity in this 
way. Interleukin-18, which is associated with 
preterm delivery and pregnancy complication [22], 
plays an important role in host defense in 
neonates. Among other effects, this cytokine 
stimulates expression of CD4 antigen, and so may 
increase the risk of HIV-1 infection. Interleukin-16 
is a chemo-attractant factor found in colostrum and 
is responsible for the traffic of leukocytes from the 
maternal circulation to the breast milk [23]. Pro-
inflammatory cytokines are essential in the 
protection of newborns from pathogens that may 
be transmitted in breast milk, especially in the first 
days of life [24], and the continued presence of IL-
12 throughout the period of measurement in this 
study supports this view. Both IL-12 and IL-18 
induce interferon-gamma (INF-gamma) secretion 
from T cells in a synergistic manner [25]. The 
presence of TGF-beta1 and TGF-beta2 may 
balance the increase of pro-inflammatory cytokines 
in breast milk [26]. Since it is not possible to 
directly modify the cytokines in human colostrum, 
we suggest that interrupting the administration of 
colostrum in this critical period of lactation may be 
advantageous.  
There is now a large amount of data that 
shows that MTCT of HIV can be dramatically 
reduced by the use of antiretroviral treatment in 
pregnancy and by the use of single dose 
treatments in the perinatal period [27]. There is 
evidence that combination prophylaxis with 
nevirapine and zidovudine may reduce the risk of 
infection even further [28]. But there is still no 
Musumeci et al - HIV load in colostrum    
28 
J Infect Developing Countries 2007; 1(1): 25-29. 
effective program, apart from avoiding 
breastfeeding, for the prevention of transmission in 
the breastfeeding period. Our data suggest single 
dose nevirapine combined with interruption of early 
feeding may reduce the risk of MTCT, since the 
high levels of both free virus, and of viral particles 
within macrophages and other mononuclear cells, 
can be avoided [29]. In this admittedly small study, 
none of the children died or acquired HIV or other 
infectious diseases.   
Acknowledgments 
The Authors are grateful to the pregnant women 
who agreed to take part to the Voluntary Counselling 
and Testing Programme and to colostrum collection, as 
well as the midwives, the nurses, and the laboratory 
technicians of the Saint Camille Medical Center, 
Ouagadougou. In particular, the skillful and patient 
collaboration of Dr. Dabogo Sia, Mme. Ouoba Thérèse, 
Mme. Ouedraogo Joséphine, Mme. Ouedraogo Louise, 
Mr. Bakamba Robert, Mme. Tiendrebeogo Agnès and 
Mme. Sanou Madomba (data entry), is gratefully 
acknowledged.  
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Corresponding Author: Salvatore Musumeci 
Department of Pharmacology, Gynecology and 
Obstetric, Pediatrics, University of Sassari, Viale San 
Pietro 43b 07100, SASSARI, Italy. Tel. 
+39/360/285505; Fax +39/095/7179690; e-mail: 
smusumeci@tiscalinet.it.  
Conflict of Interests: The authors declare that they 
have no conflict of interests.  
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Human Immunodeficiency Virus type-1 and cytokines in colostrum from HIV-infected mothers in Burkina Faso

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